Статті в журналах: "NRT1"

1

Zhang, Jingying, Zhijun Han, Yue Lu, Yanfei Zhao, Yaping Wang, Jiayue Zhang, Haoran Ma, and Yu Zhu Han. "Genome-wide identification, structural and gene expression analysis of the nitrate transporters (NRTs) family in potato (Solanum tuberosum L.)." PLOS ONE 16, no. 10 (October 21, 2021): e0257383. http://dx.doi.org/10.1371/journal.pone.0257383.

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Nitrogen (N2) is the most important source of mineral N for plant growth, which was mainly transported by nitrate transporters (NRTs). However, little is known about the NRT gene family in potato. In this study, StNRT gene family members were identified in potato. In addition, we performed StNRT subfamily classification, gene structure and distribution analysis, and conserved domain prediction using various bioinformatics tools. Totally, 39 StNRT gene members were identified in potato genome, including 33, 4 and 2 member belong to NRT1, NRT2, and NRT3, respectively. These 39 StNRT genes were randomly distributed on all chromosomes. The collinearity results show that StNRT members in potato are closely related to Solanum lycopersicum and Solanum melongena. For the expression, different members of StNRT play different roles in leaves and roots. Especially under sufficient nitrogen conditions, different members have a clear distribution in different tissues. These results provide valuable information for identifying the members of the StNRT family in potato and could provide functional characterization of StNRT genes in further research.

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2

CRISCUOLO, GIUSEPPINA, VLADIMIR TOTEV VALKOV, AURORA PARLATI, LUDOVICO MARTIN ALVES, and MAURIZIO CHIURAZZI. "Molecular characterization of the Lotus japonicus NRT1(PTR) and NRT2 families." Plant, Cell & Environment 35, no. 9 (April 19, 2012): 1567–81. http://dx.doi.org/10.1111/j.1365-3040.2012.02510.x.

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3

Newstead, Simon, and Joanne Parker. "Crystal structure of the plant nitrate transporter NRT1.1." Acta Crystallographica Section A Foundations and Advances 70, a1 (August 5, 2014): C1487. http://dx.doi.org/10.1107/s205327331408512x.

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Nitrogen uptake and assimilation is a key limiting factor for plant growth and crop productivity and also acts a major signaling molecule, controlling many aspects of plant development. Many plants obtain nitrogen through the uptake of nitrate from the soil via specific membrane transporters. Two families of nitrate transporter have been identified that act within the root cell, termed NRT1 and NRT2. NRT1 members are predominantly low affinity transporters, with KM values in the millimolar range, whereas NRT2 members are high affinity transporters, with KM values in the low micromolar range. Dual affinity transporter systems are used in biology to allow the cell to respond to changes in an external nutrient supply. In the case of nitrate transport in plants, decreasing levels of external nitrate cause an increase in the expression of NRT2 family transporter genes, in particular NRT2.1, allowing the cell to take up more of the available nitrate. However, plants have evolved a faster way of responding to nitrate levels involving post-translational control of nitrate uptake. NRT1.1 also known as CHL1, is a dual affinity nitrate transporter. In response to decreasing levels of nitrate, NRT1.1 is capable of switching between low and high KM modes, a switch achieved through the post-translational phosphorylation of an intracellular threonine. Here I will present our recently determined crystal structures of NRT1.1 in both the apo and nitrate bound forms. Together with in vitro binding and transport data we identify key residues involved in nitrate recognition and provide the first biochemical explanation for the phosphorylation controlled `dual affinity' switch observed in NRT1.1. Finally we present our model for the molecular basis of nitrate uptake via this transporter.

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4

Okamoto, Mamoru, J. John Vidmar, and Anthony D. M. Glass. "Regulation of NRT1 and NRT2 Gene Families of Arabidopsis thaliana: Responses to Nitrate Provision." Plant and Cell Physiology 44, no. 3 (March 15, 2003): 304–17. http://dx.doi.org/10.1093/pcp/pcg036.

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5

Guo, Tiancai, Hongmei Xuan, Yingying Yang, Lina Wang, Liting Wei, Yonghua Wang, and Guozhang Kang. "Transcription Analysis of Genes Encoding the Wheat Root Transporter NRT1 and NRT2 Families During Nitrogen Starvation." Journal of Plant Growth Regulation 33, no. 4 (June 19, 2014): 837–48. http://dx.doi.org/10.1007/s00344-014-9435-z.

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6

Yokoyama, Terufumi, Norio Kodama, Hitoshi Aoshima, Hanae Izu, Kazunobu Matsushita, and Mamoru Yamada. "Cloning of a cDNA for a constitutive NRT1 transporter from soybean and comparison of gene expression of soybean NRT1 transporters." Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 1518, no. 1-2 (March 2001): 79–86. http://dx.doi.org/10.1016/s0167-4781(01)00175-0.

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7

Swapnil, Prashant, Mukesh Meena, and Ashwani K. Rai. "Molecular interaction of nitrate transporter proteins with recombinant glycinebetaine results in efficient nitrate uptake in the cyanobacterium Anabaena PCC 7120." PLOS ONE 16, no. 11 (November 18, 2021): e0257870. http://dx.doi.org/10.1371/journal.pone.0257870.

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Nitrate transport in cyanobacteria is mediated by ABC-transporter, which consists of a highly conserved ATP binding cassette (ABC) and a less conserved transmembrane domain (TMD). Under salt stress, recombinant glycinebetaine (GB) not only protected the rate of nitrate transport in transgenic Anabaena PCC 7120, rather stimulated the rate by interacting with the ABC-transporter proteins. In silico analyses revealed that nrtA protein consisted of 427 amino acids, the majority of which were hydrophobic and contained a Tat (twin-arginine translocation) signal profile of 34 amino acids (1–34). The nrtC subunit of 657 amino acids contained two hydrophobic distinct domains; the N-terminal (5–228 amino acids), which was 59% identical to nrtD (the ATP-binding subunit) and the C-terminal (268–591), 28.2% identical to nrtA, suggesting C-terminal as a solute binding domain and N-terminal as ATP binding domain. Subunit nrtD consisted of 277 amino acids and its N-terminal (21–254) was an ATP binding motif. Phylogenetic analysis revealed that nitrate-ABC-transporter proteins are highly conserved among the cyanobacterial species, though variation existed in sequences resulting in several subclades. Nostoc PCC 7120 was very close to Anabaena variabilis ATCC 29413, Anabaena sp. 4–3 and Anabaena sp. CA = ATCC 33047. On the other, Nostoc spp. NIES-3756 and PCC 7524 were often found in the same subclade suggesting more work before referring it to Anabaena PCC 7120 or Nostoc PCC 7120. The molecular interaction of nitrate with nrtA was hydrophilic, while hydrophobic with nrtC and nrtD. GB interaction with nrtACD was hydrophobic and showed higher affinity compared to nitrate.

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8

Sandrock, David R., Anita N. Azarenko, Ruth M. Martin, and Nahla V. Bassil. "(271) Expression of the NRT1 Gene in Cornus and Rhododendron." HortScience 40, no. 4 (July 2005): 1020C—1020. http://dx.doi.org/10.21273/hortsci.40.4.1020c.

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The NRT1gene family encodes transport proteins with dual or low affinity for nitrate. The objectives of this experiment were to develop a system that could be used to compare the expression of the NRT1genes between species. This was accomplished by comparing sequences of NRT1homologues from various species and designing degenerate primers in regions of high homology. These primers were used to amplify a region of the NRT1gene from species of interest. A 635 bp PCR product was amplified from each species using the MD2-1 (5' ATGTTACCAAYWTGGGCMAC-3') and MD2-2 (5'-GCCAMWARCCARTAGAAAT-3') primers. The PCR products were cloned and sequenced. At the nucleotide level, CornussericeaL. `Kelseyi' and RhododendronL. `Unique' were 79.52% identical. Species-specific primers were designed and used for RT-PCR to compare NRT1expression in roots of hydroponically grown C. sericea, C. sericea `Kelseyi', and Rhododendron`Unique'. The relative levels of NRT1expression, normalized using 18S rRNA as a standard, were ≈3.2 to 1.7 to 1.0 for C. sericea, C. sericea `Kelseyi', and Rhododendron`Unique', respectively. This approach may eventually be used to examine nitrate uptake potential in different taxa of plants at different times during the growing season.

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9

Corratgé-Faillie, Claire, and Benoît Lacombe. "Substrate (un)specificity of Arabidopsis NRT1/PTR FAMILY (NPF) proteins." Journal of Experimental Botany 68, no. 12 (February 10, 2017): 3107–13. http://dx.doi.org/10.1093/jxb/erw499.

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10

Belenky, Peter A., Tiberiu G. Moga, and Charles Brenner. "Saccharomyces cerevisiae YOR071CEncodes the High Affinity Nicotinamide Riboside Transporter Nrt1." Journal of Biological Chemistry 283, no. 13 (February 6, 2008): 8075–79. http://dx.doi.org/10.1074/jbc.c800021200.

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11

Maeda, Shin-ichi, Risa Aoba, Yuma Nishino, and Tatsuo Omata. "A Novel Bacterial Nitrate Transporter Composed of Small Transmembrane Proteins." Plant and Cell Physiology 60, no. 10 (June 14, 2019): 2180–92. http://dx.doi.org/10.1093/pcp/pcz112.

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AbstractA putative silent gene of the freshwater cyanobacterium Synechococcus elongatus strain PCC 7942, encoding a small protein with two transmembrane helices, was named nrtS, since its overexpression from an inducible promoter conferred nitrate uptake activity on the nitrate transport-less NA4 mutant of S. elongatus. Homologs of nrtS, encoding proteins of 67–118 amino acid residues, are present in a limited number of eubacteria including mostly cyanobacteria and proteobacteria, but some others, e.g. the actinobacteria of the Mycobacterium tuberculosis complex, also have the gene. When expressed in NA4, the nrtS homolog of the γ-proteobacterium Marinomonas mediterranea took up nitrate with higher affinity for the substrate as compared with the S. elongatus NrtS (Km of 0.49 mM vs. 2.5 mM). Among the 61 bacterial species carrying the nrtS homolog, the marine cyanobacterium Synechococcus sp. strain PCC 7002 is unique in having two nrtS genes (nrtS1 and nrtS2) located in tandem on the chromosome. Coexpression of the two genes in NA4 resulted in nitrate uptake with a Km (NO3−) of 0.15 mM, while expression of either of the two resulted in low-affinity nitrate uptake activity with Km values of >3 mM, indicating that NrtS1 and NrtS2 form a heteromeric transporter complex. The heteromeric transporter was shown to transport nitrite as well. A Synechococcus sp. strain PCC 7002 mutant defective in the nitrate transporter (NrtP) showed a residual activity of nitrate uptake, which was ascribed to the NrtS proteins. Blue-native PAGE and immunoblotting analysis suggested a hexameric structure for the NrtS proteins.

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12

Chiba, Yasutaka, Takafumi Shimizu, Shinya Miyakawa, Yuri Kanno, Tomokazu Koshiba, Yuji Kamiya, and Mitsunori Seo. "Identification of Arabidopsis thaliana NRT1/PTR FAMILY (NPF) proteins capable of transporting plant hormones." Journal of Plant Research 128, no. 4 (March 24, 2015): 679–86. http://dx.doi.org/10.1007/s10265-015-0710-2.

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13

Migocka, M., A. Warzybok, and G. Kłobus. "The genomic organization and transcriptional pattern of genes encoding nitrate transporters 1 (NRT1) in cucumber." Plant and Soil 364, no. 1-2 (July 14, 2012): 245–60. http://dx.doi.org/10.1007/s11104-012-1345-x.

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14

Chen, Yen-Ning, and Cheng-Hsun Ho. "Concept of Fluorescent Transport Activity Biosensor for the Characterization of the Arabidopsis NPF1.3 Activity of Nitrate." Sensors 22, no. 3 (February 4, 2022): 1198. http://dx.doi.org/10.3390/s22031198.

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The NRT1/PTR FAMILY (NPF) in Arabidopsis (Arabidopsis thaliana) plays a major role as a nitrate transporter. The first nitrate transporter activity biosensor NiTrac1 converted the dual-affinity nitrate transceptor NPF6.3 into fluorescence activity sensors. To test whether this approach is transferable to other members of this family, screening for genetically encoded fluorescence transport activity sensor was performed with the member of the NPF family in Arabidopsis. In this study, NPF1.3, an uncharacterized member of NPF in Arabidopsis, was converted into a transporter activity biosensor NiTrac-NPF1.3 that responds specifically to nitrate. The emission ratio change of NiTrac-NPF1.3 triggered by the addition of nitrate reveals the important function of NPF1.3 in nitrate transport in Arabidopsis. A functional analysis of Xenopus laevis oocytes confirmed that NPF1.3 plays a role as a nitrate transporter. This new technology is applicable in plant and medical research.

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15

Zoghbi-Rodríguez, Normig M., Samuel David Gamboa-Tuz, Alejandro Pereira-Santana, Luis C. Rodríguez-Zapata, Lorenzo Felipe Sánchez-Teyer, and Ileana Echevarría-Machado. "Phylogenomic and Microsynteny Analysis Provides Evidence of Genome Arrangements of High-Affinity Nitrate Transporter Gene Families of Plants." International Journal of Molecular Sciences 22, no. 23 (December 3, 2021): 13036. http://dx.doi.org/10.3390/ijms222313036.

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Nitrate transporter 2 (NRT2) and NRT3 or nitrate-assimilation-related 2 (NAR2) proteins families form a two-component, high-affinity nitrate transport system, which is essential for the acquisition of nitrate from soils with low N availability. An extensive phylogenomic analysis across land plants for these families has not been performed. In this study, we performed a microsynteny and orthology analysis on the NRT2 and NRT3 genes families across 132 plants (Sensu lato) to decipher their evolutionary history. We identified significant differences in the number of sequences per taxonomic group and different genomic contexts within the NRT2 family that might have contributed to N acquisition by the plants. We hypothesized that the greater losses of NRT2 sequences correlate with specialized ecological adaptations, such as aquatic, epiphytic, and carnivory lifestyles. We also detected expansion on the NRT2 family in specific lineages that could be a source of key innovations for colonizing contrasting niches in N availability. Microsyntenic analysis on NRT3 family showed a deep conservation on land plants, suggesting a high evolutionary constraint to preserve their function. Our study provides novel information that could be used as guide for functional characterization of these gene families across plant lineages.

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16

Zhang, Shuaiwei, Yuepeng Zhang, Yudan Wang, Yanwei Hao, Wei Su, Guangwen Sun, Houcheng Liu, Riyuan Chen, and Shiwei Song. "Nitrogen Absorption Pattern Detection and Expression Analysis of Nitrate Transporters in Flowering Chinese Cabbage." Horticulturae 8, no. 3 (February 22, 2022): 188. http://dx.doi.org/10.3390/horticulturae8030188.

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Nitrate transporters (NRTs) play an important role in nitrate absorption and internal distribution in plant roots and other parts. Experiments were carried out to explore the sequences and expression characteristics of NRT genes, and their correlation with the N uptake in flowering Chinese cabbage. We have isolated three important BcNRTs (BcNRT1.1, BcNRT1.2, and BcNRT2.1) from flowering Chinese cabbage. Spatio-temporal expression analysis found that BcNRT1.1 and BcNRT2.1 were mainly expressed in roots, while BcNRT1.2 was more expressed in roots than in leaves during vegetative growth and was mainly expressed in leaves during reproductive growth. The NO3− uptake rate of the entire growth period was significantly correlated with BcNRT1.1 and BcNRT1.2 expression in roots. In addition, the total N content was increased with the increase in NO3− concentration in flowering Chinese cabbage. The NH4+ uptake was slightly induced by NH4+, but the total N content had no significant difference under the NH4+ concentration of 1–8 mmol/L. We also found that lower concentrations of NH4+ promoted the expression of BcNRT1.1 and BcNRT1.2 while inhibiting the expression of BcNRT2.1 in the roots of flowering Chinese cabbage. The amount of total N uptake in the treatment with 25/75 of NH4+/NO3− was significantly higher than that of the other two treatments (0/100 and 50/50). In the mixture of NH4+ and NO3−, total N uptake was significantly correlated with the BcNRT1.2 expression. We concluded that mixed nutrition with an NH4+/NO3− of 25/75 could significantly increase total nitrogen uptake in flowering Chinese cabbage, in which two members of the NRT1 subfamily (BcNRT1.1 and BcNRT1.2) might play a major regulatory role in it. This study is a beneficial attempt to dig deeper into the NRT genes resources and lays the foundation for the ultimate use of genetic improvement methods to increase the NUE with less nitrogen fertilizer in flowering Chinese cabbage.

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17

Belenky, Peter, Rebecca Stebbins, Katrina L. Bogan, Charles R. Evans, and Charles Brenner. "Nrt1 and Tna1-Independent Export of NAD+ Precursor Vitamins Promotes NAD+ Homeostasis and Allows Engineering of Vitamin Production." PLoS ONE 6, no. 5 (May 11, 2011): e19710. http://dx.doi.org/10.1371/journal.pone.0019710.

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18

Weichert, Annett, Christopher Brinkmann, Nataliya Y. Komarova, Daniela Dietrich, Kathrin Thor, Stefan Meier, Marianne Suter Grotemeyer, and Doris Rentsch. "AtPTR4 and AtPTR6 are differentially expressed, tonoplast-localized members of the peptide transporter/nitrate transporter 1 (PTR/NRT1) family." Planta 235, no. 2 (September 9, 2011): 311–23. http://dx.doi.org/10.1007/s00425-011-1508-7.

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19

Huang, Wan-Ru, Pin-Yi Liu, Jie Hsu, Xiuzhen Li, and Liping Deng. "Assessment of Near-Real-Time Satellite Precipitation Products from GSMaP in Monitoring Rainfall Variations over Taiwan." Remote Sensing 13, no. 2 (January 8, 2021): 202. http://dx.doi.org/10.3390/rs13020202.

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This study assessed four near-real-time satellite precipitation products (NRT SPPs) of Global Satellite Mapping of Precipitation (GSMaP)—NRT v6 (hereafter NRT6), NRT v7 (hereafter NRT7), Gauge-NRT v6 (hereafter GNRT6), and Gauge-NRT v7 (hereafter GNRT7)— in representing the daily and monthly rainfall variations over Taiwan, an island with complex terrain. The GNRT products are the gauge-adjusted version of NRT products. Evaluations for warm (May–October) and cold months (November–April) were conducted from May 2017 to April 2020. By using observations from more than 400 surface gauges in Taiwan as a reference, our evaluations showed that GNRT products had a greater error than NRT products in underestimating the monthly mean rainfall, especially during the warm months. Among SPPs, NRT7 performed best in quantitative monthly mean rainfall estimation; however, when examining the daily scale, GNRT6 and GNRT7 were superior, particularly for monitoring stronger (i.e., more intense) rainfall events during warm and cold months, respectively. Spatially, the major improvement from NRT6 to GNRT6 (from NRT7 to GNRT7) in monitoring stronger rainfall events over southwestern Taiwan was revealed during warm (cold) months. From NRT6 to NRT7, the improvement in daily rainfall estimation primarily occurred over southwestern and northwestern Taiwan during the warm and cold months, respectively. Possible explanations for the differences between the ability of SPPs are attributed to the algorithms used in SPPs. These findings highlight that different NRT SPPs of GSMaP should be used for studying or monitoring the rainfall variations over Taiwan for different purposes (e.g., warning of floods in different seasons, studying monthly or daily precipitation features in different seasons, etc.).

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20

Huang, Wan-Ru, Pin-Yi Liu, Jie Hsu, Xiuzhen Li, and Liping Deng. "Assessment of Near-Real-Time Satellite Precipitation Products from GSMaP in Monitoring Rainfall Variations over Taiwan." Remote Sensing 13, no. 2 (January 8, 2021): 202. http://dx.doi.org/10.3390/rs13020202.

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This study assessed four near-real-time satellite precipitation products (NRT SPPs) of Global Satellite Mapping of Precipitation (GSMaP)—NRT v6 (hereafter NRT6), NRT v7 (hereafter NRT7), Gauge-NRT v6 (hereafter GNRT6), and Gauge-NRT v7 (hereafter GNRT7)— in representing the daily and monthly rainfall variations over Taiwan, an island with complex terrain. The GNRT products are the gauge-adjusted version of NRT products. Evaluations for warm (May–October) and cold months (November–April) were conducted from May 2017 to April 2020. By using observations from more than 400 surface gauges in Taiwan as a reference, our evaluations showed that GNRT products had a greater error than NRT products in underestimating the monthly mean rainfall, especially during the warm months. Among SPPs, NRT7 performed best in quantitative monthly mean rainfall estimation; however, when examining the daily scale, GNRT6 and GNRT7 were superior, particularly for monitoring stronger (i.e., more intense) rainfall events during warm and cold months, respectively. Spatially, the major improvement from NRT6 to GNRT6 (from NRT7 to GNRT7) in monitoring stronger rainfall events over southwestern Taiwan was revealed during warm (cold) months. From NRT6 to NRT7, the improvement in daily rainfall estimation primarily occurred over southwestern and northwestern Taiwan during the warm and cold months, respectively. Possible explanations for the differences between the ability of SPPs are attributed to the algorithms used in SPPs. These findings highlight that different NRT SPPs of GSMaP should be used for studying or monitoring the rainfall variations over Taiwan for different purposes (e.g., warning of floods in different seasons, studying monthly or daily precipitation features in different seasons, etc.).

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21

Zhang, Hao, Shuang Li, Mengyao Shi, Sheliang Wang, Lei Shi, Fangsen Xu, and Guangda Ding. "Genome-Wide Systematic Characterization of the NPF Family Genes and Their Transcriptional Responses to Multiple Nutrient Stresses in Allotetraploid Rapeseed." International Journal of Molecular Sciences 21, no. 17 (August 19, 2020): 5947. http://dx.doi.org/10.3390/ijms21175947.

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NITRATE TRANSPORTER 1 (NRT1)/PEPTIDE TRANSPORTER (PTR) family (NPF) proteins can transport various substrates, and play crucial roles in governing plant nitrogen (N) uptake and distribution. However, little is known about the NPF genes in Brassica napus. Here, a comprehensive genome-wide systematic characterization of the NPF family led to the identification of 193 NPF genes in the whole genome of B. napus. The BnaNPF family exhibited high levels of genetic diversity among sub-families but this was conserved within each subfamily. Whole-genome duplication and segmental duplication played a major role in BnaNPF evolution. The expression analysis indicated that a broad range of expression patterns for individual gene occurred in response to multiple nutrient stresses, including N, phosphorus (P) and potassium (K) deficiencies, as well as ammonium toxicity. Furthermore, 10 core BnaNPF genes in response to N stress were identified. These genes contained 6–13 transmembrane domains, located in plasma membrane, that respond discrepantly to N deficiency in different tissues. Robust cis-regulatory elements were identified within the promoter regions of the core genes. Taken together, our results suggest that BnaNPFs are versatile transporters that might evolve new functions in B. napus. Our findings benefit future research on this gene family.

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22

He, Yingdui, Ruimei Li, Fei Lin, Ying Xiong, Lixia Wang, Bizun Wang, Jianchun Guo, and Chengxiao Hu. "Transcriptome Changes Induced by Different Potassium Levels in Banana Roots." Plants 9, no. 1 (December 19, 2019): 11. http://dx.doi.org/10.3390/plants9010011.

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Potassium plays an important role in enhancing plant resistance to biological and abiotic stresses and improving fruit quality. To study the effect of potassium nutrient levels on banana root growth and its regulation mechanism, four potassium concentrations were designed to treat banana roots from no potassium to high potassium. The results indicated that K2 (3 mmol/L K2SO4) treatment was a relatively normal potassium concentration for the growth of banana root, and too high or too low potassium concentration was not conducive to the growth of banana root. By comparing the transcriptome data in each treatment in pairs, 4454 differentially expressed genes were obtained. There were obvious differences in gene function enrichment in root systems treated with different concentrations of potassium. Six significant expression profiles (profile 0, 1, 2, 7, 9 and 13) were identified by STEM analysis. The hub genes were FKF1, HsP70-1, NRT1/PTR5, CRY1, and ZIP11 in the profile 0; CYP51 in profile 1; SOS1 in profile 7; THA, LKR/SDH, MCC, C4H, CHI, F3′H, 2 PR1s, BSP, TLP, ICS, RO, chitinase and peroxidase in profile 9. Our results provide a comprehensive and systematic analysis of the gene regulation network in banana roots under different potassium stress.

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23

Guo, Hao, Xuyou He, Hao Zhang, Ronglei Tan, Jinpeng Yang, Fangsen Xu, Sheliang Wang, Chunlei Yang, and Guangda Ding. "Physiological Responses of Cigar Tobacco Crop to Nitrogen Deficiency and Genome-Wide Characterization of the NtNPF Family Genes." Plants 11, no. 22 (November 11, 2022): 3064. http://dx.doi.org/10.3390/plants11223064.

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Tobacco prefers nitrate as a nitrogen (N) source. However, little is known about the molecular components responsible for nitrate uptake and the physiological responses of cigar tobacco to N deficiency. In this study, a total of 117 nitrate transporter 1 (NRT1) and peptide transporter (PTR) family (NPF) genes were comprehensively identified and systematically characterized in the whole tobacco genome. The NtNPF members showed significant genetic diversity within and across subfamilies but showed conservation between subfamilies. The NtNPF genes are dispersed unevenly across the chromosomes. The phylogenetic analysis revealed that eight subfamilies of NtNPF genes are tightly grouped with their orthologues in Arabidopsis. The promoter regions of the NtNPF genes had extensive cis-regulatory elements. Twelve core NtNPF genes, which were strongly induced by N limitation, were identified based on the RNA-seq data. Furthermore, N deprivation severely impaired plant growth of two cigar tobaccos, and CX26 may be more sensitive to N deficiency than CX14. Moreover, 12 hub genes respond differently to N deficiency between the two cultivars, indicating the vital roles in regulating N uptake and transport in cigar tobacco. The findings here contribute towards a better knowledge of the NtNPF genes and lay the foundation for further functional analysis of cigar tobacco.

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24

Wang, Yongxin, Kang Wei, Li Ruan, Peixian Bai, Liyun Wu, Liyuan Wang, and Hao Cheng. "Systematic Investigation and Expression Profiles of the Nitrate Transporter 1/Peptide Transporter Family (NPF) in Tea Plant (Camellia sinensis)." International Journal of Molecular Sciences 23, no. 12 (June 15, 2022): 6663. http://dx.doi.org/10.3390/ijms23126663.

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NRT1/PTR FAMILY (NPF) genes are characterized as nitrate and peptide transporters that played important roles in various substrates transport in plants. However, little is known about the NPF gene in tea plants. Here, a total of 109 CsNPF members were identified from the tea plant genome, and divided into 8 groups according to their sequence characteristics and phylogenetic relationship. Gene structure and conserved motif analysis supported the evolutionary conservation of CsNPFs. Many hormone and stress response cis-acting elements and transcription factor binding sites were found in CsNPF promoters. Syntenic analysis suggested that multiple duplication types contributed to the expansion of NPF gene family in tea plants. Selection pressure analysis showed that CsNPF genes experienced strong purifying selective during the evolution process. The distribution of NPF family genes revealed that 8 NPF subfamilies were formed before the divergence of eudicots and monocots. Transcriptome analysis showed that CsNPFs were expressed differently in different tissues of the tea plant. The expression of 20 CsNPF genes at different nitrate concentrations was analyzed, and most of those genes responded to nitrate resupply. Subcellular localization showed that both CsNPF2.3 and CsNPF6.1 were localized in the plasma membrane, which was consistent with the characteristics of transmembrane proteins involved in NO3- transport. This study provides a theoretical basis for further investigating the evolution and function of NPF genes.

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25

Zhang, Yinkun, Yanling Yin, Erick Amombo, Xiaoning Li, and Jinmin Fu. "Different mowing frequencies affect nutritive value and recovery potential of forage bermudagrass." Crop and Pasture Science 71, no. 6 (2020): 610. http://dx.doi.org/10.1071/cp19369.

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Mowing is one of the most effective methods of pasture management, and frequency plays a critical role in management strategies. Bermudagrass (Cynodon dactylon (L.) Pers.) is a highly valuable forage grass due to its exceptionally high mowing recovery rate and its high potential to be used as a forage crop. In China, bermudagrass is increasingly becoming a crucial forage crop because of the growing demand from the livestock industry. The objective of the present study was to determine the effect of mowing frequencies on forage bermudagrass (‘Wrangler’) yields and nutritive value. Four treatments with different mowing frequencies (2, 4, 6 and 12 weeks) were evaluated. Harvested grasses were assessed for yield and nutritive value. The shoot dry weight, crude fibre and N content did not exhibit any difference at various mowing frequencies. The highest content of crude protein was attained at the 2-weeks mowing frequency, although the 4-weeks mowing frequency resulted in a relatively higher shoot fresh weight, crude fat content, water content, P concentration and plant height. A persistently high upregulation of NRT1, PHT1, PHT2, AOC, AOS, MYC2 and NCED1 genes were observed at 4-weeks frequency. Yield was highest at 4- and 6-weeks mowing frequencies. Consequently, the 4-weeks frequency was considered to be the optimal mowing frequency in view of the forage quality and regrowth capacity.

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Lisker, Antonia, Andreas Maurer, Thomas Schmutzer, Ebrahim Kazman, Hilmar Cöster, Josef Holzapfel, Erhard Ebmeyer, Ahmad M. Alqudah, Wiebke Sannemann, and Klaus Pillen. "A Haplotype-Based GWAS Identified Trait-Improving QTL Alleles Controlling Agronomic Traits under Contrasting Nitrogen Fertilization Treatments in the MAGIC Wheat Population WM-800." Plants 11, no. 24 (December 14, 2022): 3508. http://dx.doi.org/10.3390/plants11243508.

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The multi-parent-advanced-generation-intercross (MAGIC) population WM-800 was developed by intercrossing eight modern winter wheat cultivars to enhance the genetic diversity present in breeding populations. We cultivated WM-800 during two seasons in seven environments under two contrasting nitrogen fertilization treatments. WM-800 lines exhibited highly significant differences between treatments, as well as high heritabilities among the seven agronomic traits studied. The highest-yielding WM-line achieved an average yield increase of 4.40 dt/ha (5.2%) compared to the best founder cultivar Tobak. The subsequent genome-wide-association-study (GWAS), which was based on haplotypes, located QTL for seven agronomic traits including grain yield. In total, 40, 51, and 46 QTL were detected under low, high, and across nitrogen treatments, respectively. For example, the effect of QYLD_3A could be associated with the haplotype allele of cultivar Julius increasing yield by an average of 4.47 dt/ha (5.2%). A novel QTL on chromosome 2B exhibited pleiotropic effects, acting simultaneously on three-grain yield components (ears-per-square-meter, grains-per-ear, and thousand-grain-weight) and plant-height. These effects may be explained by a member of the nitrate-transporter-1 (NRT1)/peptide-family, TaNPF5.34, located 1.05 Mb apart. The WM-800 lines and favorable QTL haplotypes, associated with yield improvements, are currently implemented in wheat breeding programs to develop advanced nitrogen-use efficient wheat cultivars.

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Rani, Vaishali, and Gergely Maróti. "Light-Dependent Nitrate Removal Capacity of Green Microalgae." International Journal of Molecular Sciences 24, no. 1 (December 21, 2022): 77. http://dx.doi.org/10.3390/ijms24010077.

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In the present study, Chlamydomonas sp. MACC-216 was used to investigate total nitrate removal in TAP medium with sodium nitrate as the sole nitrogen source under several light conditions made up of permuted combinations of three light colors (referred to as blue, red, and white light) and three light intensities (50 µmol m−2 s−1, 100 µmol m−2 s−1, and 250 µmol m−2 s−1). It was observed that nitrate removal efficiency is influenced by light color as well as light intensity. Additionally, Chlamydomonas sp. MACC-216 was cultivated in synthetic wastewater under four light conditions, namely, Blue 250, Blue 125 + Red 125, Red 250, and White 250, where it showed the highest nitrate removal efficiency and nitrate reductase activity under the Blue 125 + Red 125 light condition. To observe the impact of light color on the nitrate removal capacity of Chlamydomonas sp. MACC-216, the expression of five genes participating in nitrate transport and reduction (NRT1, NRT2.1. NRT2.2, NIA, and MCP) was also analyzed; these genes showed the highest expression under the Blue 125 + Red 125 light condition. Based on the above-mentioned findings, the blue + red light combination emerged as a promising light combination for nitrate removal. Hence, our study suggests the importance of the blue + red light combination together with high light intensity, as the optimal light condition for nitrate removal from synthetic wastewater in comparison to other monochromatic lights with high light intensity.

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Sakamoto, Toshio, Kaori Inoue-Sakamoto, and Donald A. Bryant. "A Novel Nitrate/Nitrite Permease in the Marine CyanobacteriumSynechococcus sp. Strain PCC 7002." Journal of Bacteriology 181, no. 23 (December 1, 1999): 7363–72. http://dx.doi.org/10.1128/jb.181.23.7363-7372.1999.

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ABSTRACT The nrtP and narB genes, encoding nitrate/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-type nitrate transporters found in diverse organisms. An nrtP mutant strain consumes nitrate at a 4.5-fold-lower rate than the wild type, and this mutant grew exponentially on a medium containing 12 mM nitrate at a rate approximately 2-fold lower than that of the wild type. The nrtP mutant cells could not consume nitrite as rapidly as the wild type at pH 10, suggesting that NrtP also functions in nitrite uptake. A narB mutant was unable to grow on a medium containing nitrate as a nitrogen source, although this mutant could grow on media containing urea or nitrite with rates similar to those of the wild type. Exogenously added nitrite enhanced the in vivo activity of nitrite reductase in the narBmutant; this suggests that nitrite acts as a positive effector of nitrite reductase. Transcripts of the nrtP andnarB genes were detected in cells grown on nitrate but were not detected in cells grown on urea or ammonia. Transcription of thenrtP and narB genes is probably controlled by the NtcA transcription factor for global nitrogen control. The discovery of a nitrate/nitrite permease in Synechococcussp. strain PCC 7002 suggests that significant differences in nutrient transporters may occur in marine and freshwater cyanobacteria.

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Wang, Xiaoli, Xiaofeng Cai, Chenxi Xu, and Quanhua Wang. "Identification and characterization of the NPF, NRT2 and NRT3 in spinach." Plant Physiology and Biochemistry 158 (January 2021): 297–307. http://dx.doi.org/10.1016/j.plaphy.2020.11.017.

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Aslam, Ali, Shengjie Zhao, Muhammad Azam, Xuqiang Lu, Nan He, Bingbing Li, Junling Dou, Hongju Zhu, and Wenge Liu. "Comparative analysis of primary metabolites and transcriptome changes between ungrafted and pumpkin-grafted watermelon during fruit development." PeerJ 8 (January 6, 2020): e8259. http://dx.doi.org/10.7717/peerj.8259.

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Grafting has been reported as a factor that influences fruit quality. However, a comprehensive study of the metabolic profile related to fruit quality and the underlying molecular mechanism in grafted watermelon has not been carried out. Metabolomics and transcriptome analysis were performed on both pumpkin-grafted watermelon and ungrafted watermelon at different developmental stages. In total, 56 primary metabolites were identified with either high or low abundance between ungrafted and pumpkin-grafted watermelon. The results indicated that ornithine, arginine, lysine (amino acids), glucose, sucrose, glucosamine (sugars), malic acid, fumaric acid and succinic acid (organic acids) were among the dominant metabolites influencing fruit quality. Additionally, comparative RNA sequence analysis on grafted and ungrafted watermelon yielded 729, 174, 128 and 356 differentially expressed genes at 10, 18, 26 and 34 days after pollination (DAP), respectively. Functional annotations of these genes indicated that grafting significantly altered the biological and metabolic processes related to fruit quality. Our comparative metabolomics and transcriptome analysis revealed that FBA2, FK, SuSy, SPS, IAI, AI and sugar transporter gene (SWT3b) might play a central role in the accumulation of glucose and sucrose, whereas higher malic acid content was attributed to high down regulation of ALMT13 and ALMT8 in pumpkin-grafted watermelon. Changes in the ornithine, glutamine, alanine, tyrosine, valine, asparagine, phenylalanine, arginine and tryptophan contents were consistent with the transcript level of their metabolic genes such as NAOD, GS, AGT, TaT, aDH1, OGDH, aDC, 4CL 1, PaL, CaT and two nitrate transporter genes (NRT1) in pumpkin-grafted watermelon. This study provides the basis for understanding the graft-responsive changes in the metabolic profile and regulatory mechanism related to fruit quality.

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Tarroum, Mohamed, Walid Ben Romdhane, Ahmed Abdelrahim Mohamed Ali, Fahad Al-Qurainy, Abdullah Al-Doss, Lotfi Fki, and Afif Hassairi. "Harnessing the Rhizosphere of the Halophyte Grass Aeluropus littoralis for Halophilic Plant-Growth-Promoting Fungi and Evaluation of Their Biostimulant Activities." Plants 10, no. 4 (April 16, 2021): 784. http://dx.doi.org/10.3390/plants10040784.

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Hydroponic systems have gained interest and are increasingly used in hot and dry desert areas. Numbers of benefits are offered by hydroponic systems such as the ability to save water, enhance nutrients use efficiency, easy environmental control, and prevention of soil-borne diseases. However, the high consumption of chemical fertilizers for nutrient solution and the sensitivity of closed hydroponic systems to salinity are issues that need solutions. Thus, the main goal of our research activities is to isolate plant growth promoting fungi in order to develop sustainable hydroponic systems. We are working on isolating and testing the possibility to incorporate the cell-free filtrate (CFF) of plant growth promoting fungi (PGPF) in the composition of the nutrient solution. In this work, we isolated six strains of PGPF from the rhizosphere of the halophyte grass Aeluropus littoralis. Phylogenetic analyses of DNA sequences amplified by ITS1 and ITS4 primers identified the isolated fungi as: Byssochlamys spectabilis, Chaetomium globosum, Cephalotheca foveolata, Penicillium melinii, Alternaria tenuissima, and Nigrospora chinensis. The promoting of vigor in tobacco seedlings was used as criteria to evaluate the biostimulant activity of these fungi by adding either their mycelia (DE: direct effect) or their cell-free filtrates (CFF: indirect effect) to the plant-growth media. The best significant growth stimulation was obtained with plants treated by B. spectabilis. However, only the CFFs of Byssochlamys spectabilis (A5.1) and Penicillium melinii (A8) when added at a dilution factor of 1/50 to half-strength nutritive solution (0.5NS) resulted in significant improvement of all assessed growth parameters. Indeed, the A5.1CFF and A8CFF in 0.5NS induced a significant better increase in the biomass production when compared to NS or 0.5NS alone. All fungi produced indole acetic acid in the CFFs, which could be one of the key factors explaining their biostimulant activities. Furthermore, six genes involved in nitrogen-metabolism (NR1 and NRT1), auxin biosynthesis (Tryp1 and YUCCA6-like), and brassinosteroid biosynthesis (DET2 and DWF4) were shown to be induced in roots or leaves following treatment of plants with the all CFFs. This work opens up a prospect to study in deep the biostimulant activity of PGPFs and their applications to decrease the requirement of chemical fertilizers in the hydroponic growing systems.

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Akhtar, Naureen, Eugenia Karabika, Duncan A. Rouch, James R. Kinghorn, Anthony D. M. Glass, and Shiela E. Unkles. "High-affinity nitrate/nitrite transporters NrtA and NrtB of Aspergillus nidulans exhibit high specificity and different inhibitor sensitivity." Microbiology 161, no. 7 (July 1, 2015): 1435–46. http://dx.doi.org/10.1099/mic.0.000088.

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Zhao, Zeqi, Mengdi Li, Weiwei Xu, Ji-Hong Liu, and Chunlong Li. "Genome-Wide Identification of NRT Gene Family and Expression Analysis of Nitrate Transporters in Response to Salt Stress in Poncirus trifoliata." Genes 13, no. 7 (June 22, 2022): 1115. http://dx.doi.org/10.3390/genes13071115.

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The uptake and transportation of nitrate play a crucial role in plant growth and development. These processes mostly depend on nitrate transporters (NRT), which guarantee the supplement of nutrition in the plant. In this study, genes encoding NRT with Major Facilitator Superfamily (MFS) domain were identified in trifoliate orange (Poncirus trifoliata (L.) Raf.). Totally, 56 NRT1s, 6 NRT2s, and 2 NAR2s were explored. The bioinformation analysis, including protein characteristics, conserved domain, motif, phylogenetic relationship, cis-acting element, and synteny correlation, indicated the evolutionary conservation and functional diversity of NRT genes. Additionally, expression profiles of PtrNRTs in different tissues demonstrated that NRT genes possessed spatio-temporal expression specificity. Further, the salt condition was certified to induce the expression of some NRT members, like PtrNPF2.1, PtrNPF7.4, and PtrNAR2.1, proposing the potential role of these NRTs in salt stress response. The identification of NRT genes and the expression pattern analysis in various tissues and salt stress lay a foundation for future research between nitrogen transport and salt resistance in P. trifoliata.

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Wang, Shuai, Hao Cheng, Linlin Wang, Rui Zhao, and Dawei Guan. "Overexpression of NRF1-742 or NRF1-772 Reduces Arsenic-Induced Cytotoxicity and Apoptosis in Human HaCaT Keratinocytes." International Journal of Molecular Sciences 21, no. 6 (March 16, 2020): 2014. http://dx.doi.org/10.3390/ijms21062014.

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Increasing evidence indicates that human exposure to inorganic arsenic causes cutaneous diseases and skin cancers. Nuclear factor erythroid 2-like 1 (NRF1) belongs to the cap “n” collar (CNC) basic-region leucine zipper (bZIP) transcription factor family and regulates antioxidant response element (ARE) genes. The human NRF1 gene is transcribed into multiple isoforms, which contain 584, 616, 742, 761, or 772 amino acids. We previously demonstrated that the long isoforms of NRF1 (i.e., NRF1-742, NRF1-761 and NRF1-772) are involved in the protection of human keratinocytes from acute arsenic cytotoxicity by enhancing the cellular antioxidant response. The aim of the current study was to investigate the roles of NRF1-742 and NRF1-772 in the arsenic-induced antioxidant response and cytotoxicity. We found that overexpression of NRF1-742 or NRF1-772 in human HaCaT keratinocytes decreased susceptibility to arsenic-induced apoptosis and cytotoxicity. In addition, we characterized the different protein bands observed for NRF1-742 and NRF1-772 by western blotting. The posttranslational modifications and nuclear translocation of these isoforms differed and were partially affected by arsenic exposure. Antioxidant protein levels were increased in the NRF1-742 and NRF1-772-overexpressing cell lines. The upregulation of antioxidant protein levels was partly due to the translation of nuclear factor erythroid 2-like 2 (NRF2) and its increased nuclear transport. Overall, overexpression of NRF1-742 and NRF1-772 protected HaCaT cells from arsenic-induced cytotoxicity, mainly through translational modifications and the promotion of antioxidant gene expression.

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Chuckran, Christopher A., Anthony R. Cillo, Jessica Moskovitz, Abigail E. Overacre-Delgoffe, Ashwin Somasundaram, John M. Kirkwood, James Luketich, et al. "Neuropilin-1 Amplifies Regulatory T cell Function in Human Cancer." Journal of Immunology 204, no. 1_Supplement (May 1, 2020): 244.2. http://dx.doi.org/10.4049/jimmunol.204.supp.244.2.

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Abstract Regulatory T cells (Tregs) maintain peripheral tolerance but also contribute to cancer progression by dampening anti-tumor immunity. Knocking out neuropilin-1 (NRP1) on murine Tregs attenuates tumor growth by rendering intratumoral Tregs non-suppressive. Whereas NRP1 is constitutively expressed on all mouse Tregs, NRP1 is infrequently expressed on Tregs in the peripheral blood of healthy humans. We hypothesized that (1) NRP1 expression is upregulated in cancer patients, (2) NRP1 marks highly suppressive human Tregs, and (3) sustained Treg activation initiates NRP1 expression. NRP1+ Tregs are enriched in tumors across numerous cancer types but largely devoid in non-cancerous inflamed and healthy tissues. Intratumoral NRP1 expression negatively correlates with disease-free survival in head and neck squamous cell carcinoma (HNSCC; p = 0.03; Hazard ratio = 5.4). Intratumoral NRP1+ Tregs upregulate a suppressive module indicated by co-expression of TIGIT, CCR8, and multiple TNF receptor super family members. Correspondingly, NRP1+ Tregs potently suppress CD8+ T cell proliferation in vitro, which can be disrupted with NRP1-specific blockade. Furthermore, Treg activation in vitro drives NRP1 expression, and sustained TCR signals are required to maintain NRP1 expression. We conclude that NRP1+ Tregs are selectively enriched in human cancer and oppose anti-tumor immunity. Therefore, destabilizing intratumoral Tregs using NRP1 blockade may complement other T cell therapies such as anti-PD1 blockade.

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Navarro-González, Carmen, Alba Huerga-Gómez, and Pietro Fazzari. "Nrg1 Intracellular Signaling Is Neuroprotective upon Stroke." Oxidative Medicine and Cellular Longevity 2019 (September 8, 2019): 1–15. http://dx.doi.org/10.1155/2019/3930186.

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The schizophrenia risk gene NRG1 controls the formation of excitatory and inhibitory synapses in cortical circuits. While the expression of different NRG1 isoforms occurs during development, adult neurons primarily express the CRD-NRG1 isoform characterized by a highly conserved intracellular domain (NRG1-ICD). We and others have demonstrated that Nrg1 intracellular signaling promotes dendrite elongation and excitatory connections during neuronal development. However, the role of Nrg1 intracellular signaling in adult neurons and pathological conditions remains largely unaddressed. Here, we investigated the role of Nrg1 intracellular signaling in neuroprotection and stroke. Our bioinformatic analysis revealed the evolutionary conservation of the NRG1-ICD and a decrease in NRG1 expression with age in the human frontal cortex. Hence, we first evaluated whether Nrg1 signaling may affect pathological hallmarks in an in vitro model of neuronal senescence; however, our data failed to reveal a role for Nrg1 in the activation of the stress-related pathway p38 MAPK and DNA damage. Previous studies demonstrated that the soluble EGF domain of Nrg1 alleviated brain ischemia, a pathological process involving the generation of free radicals, reactive oxygen species (ROS), and excitotoxicity. Hence, we tested the hypothesis that Nrg1 intracellular signaling could be neuroprotective in stroke. We discovered that Nrg1 expression significantly increased neuronal survival upon oxygen-glucose deprivation (OGD), an established in vitro model for stroke. Notably, the specific activation of Nrg1 intracellular signaling by expression of the Nrg1-ICD protected neurons from OGD. Additionally, time-lapse experiments confirmed that Nrg1 intracellular signaling increased the survival of neurons exposed to OGD. Finally, we investigated the relevance of Nrg1 intracellular signaling in stroke in vivo. Using viral vectors, we expressed the Nrg1-ICD in cortical neurons and subsequently challenged them by a focal hemorrhagic stroke; our data indicated that Nrg1 intracellular signaling improved neuronal survival in the infarcted area. Altogether, these data highlight Nrg1 intracellular signaling as neuroprotective upon ischemic lesion both in vitro and in vivo. Given the complexity of the neurotoxic effects of stroke and the involvement of various mechanisms, such as the generation of ROS, excitotoxicity, and inflammation, further studies are required to determine the molecular bases of the neuroprotective effect of Nrg1 intracellular signaling. In conclusion, our research highlights the stimulation of Nrg1 intracellular signaling as a promising target for cortical stroke treatment.

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Miao, Hua-Quan, Shay Soker, Leonard Feiner, José Luis Alonso, Jonathan A. Raper, and Michael Klagsbrun. "Neuropilin-1 Mediates Collapsin-1/Semaphorin III Inhibition of Endothelial Cell Motility." Journal of Cell Biology 146, no. 1 (July 12, 1999): 233–42. http://dx.doi.org/10.1083/jcb.146.1.233.

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Neuropilin-1 (NRP1) is a receptor for two unrelated ligands with disparate activities, vascular endothelial growth factor-165 (VEGF165), an angiogenesis factor, and semaphorin/collapsins, mediators of neuronal guidance. To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined. Collapsin-1 inhibited the motility of porcine aortic EC (PAEC) expressing NRP1 alone; coexpressing KDR and NRP1 (PAEC/KDR/NRP1), but not parental PAEC; or PAEC expressing KDR alone. The motility of PAEC expressing NRP1 was inhibited by 65–75% and this inhibition was abrogated by anti-NRP1 antibody. In contrast, VEGF165 stimulated the motility of PAEC/KDR/NRP1. When VEGF165 and collapsin-1 were added simultaneously to PAEC/KDR/NRP1, dorsal root ganglia (DRG), and COS-7/NRP1 cells, they competed with each other in EC motility, DRG collapse, and NRP1-binding assays, respectively, suggesting that the two ligands have overlapping NRP1 binding sites. Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner. In an in vitro angiogenesis assay, collapsin-1 inhibited the capillary sprouting of EC from rat aortic ring segments. These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

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Zhang, Yan, Jin Su, Yue Teng, Jian Zhang, Jiang Wang, Kun Li, Libo Yao, and Xu Li. "Nrp1, a Neuronal Regulator, Enhances DDR2-ERK-Runx2 Cascade in Osteoblast Differentiation via Suppression of DDR2 Degradation." Cellular Physiology and Biochemistry 36, no. 1 (2015): 75–84. http://dx.doi.org/10.1159/000374054.

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Background: Osteoblastogenesis is under delicate control by multiple factors and hormones. Recent reports indicated the involvement of immunological and neuronal regulators. However, the role of neuropilin 1 (Nrp1) in osteoblastogenesis remains obscure. Methods: Real-time PCR was carried out to detect the mRNA of osteoblastic markers, Nrp1, and discoidin domain receptor 2 (DDR2). Immunoblot was performed to test the protein of Nrp1 and DDR2. Osteogenic differentiation was evaluated by mRNA analysis of osteogenic markers, and determination of ALP activity and OCN secretion. The intercellular signaling effectors were examined by immunoblot. Immunofluorescent assays were performed to detect the localization of Nrp1 and DDR2. Half-life determination assay was executed to test the DDR2 stability. Results: The expression of Nrp1 paralleled with that of DDR2 during osteoblastogensis. Nrp1 overexpression enhanced DDR2-induced stimulation of osteoblastogensis, whereas Nrp1 silencing caused attenuation. Nrp1 overexpression increased the phosphorylation of DDR2, ERK1/2 and Runx2. Nrp1 co-localized with DDR2 in the cellular membrane of differentiated MC3T3-E1. Enhanced or attenuated Nrp1 expression did not alter the mRNA transcript of DDR2. Nrp1 overexpression prolonged the half-life of DDR2 protein. Conclusion: Our results originally demonstrated the stimulatory role of Nrp1 in DDR2-induced osteoblast differentiation, providing molecular evidence for exploiting Nrp1 and DDR2 as targets to treat bone-related disease.

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Werr, Lisa, Dennis Plenker, Marcel A. Dammert, Carina Lorenz, Johannes Brägelmann, Hannah L. Tumbrink, Sebastian Klein, et al. "CD74-NRG1 Fusions Are Oncogenic In Vivo and Induce Therapeutically Tractable ERBB2:ERBB3 Heterodimerization." Molecular Cancer Therapeutics 21, no. 5 (February 28, 2022): 821–30. http://dx.doi.org/10.1158/1535-7163.mct-21-0820.

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Abstract NRG1 fusions are recurrent somatic genome alterations occurring across several tumor types, including invasive mucinous lung adenocarcinomas and pancreatic ductal adenocarcinomas and are potentially actionable genetic alterations in these cancers. We initially discovered CD74-NRG1 as the first NRG1 fusion in lung adenocarcinomas, and many additional fusion partners have since been identified. Here, we present the first CD74-NRG1 transgenic mouse model and provide evidence that ubiquitous expression of the CD74-NRG1 fusion protein in vivo leads to tumor development at high frequency. Furthermore, we show that ERBB2:ERBB3 heterodimerization is a mechanistic event in transformation by CD74-NRG1 binding physically to ERBB3 and that CD74-NRG1–expressing cells proliferate independent of supplemented NRG1 ligand. Thus, NRG1 gene fusions are recurrent driver oncogenes that cause oncogene dependency. Consistent with these findings, patients with NRG1 fusion-positive cancers respond to therapy targeting the ERBB2:ERBB3 receptors.

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Mikolaskova, Barbora, Matus Jurcik, Ingrid Cipakova, Tomas Selicky, Jan Jurcik, Silvia Bagelova Polakova, Erika Stupenova, et al. "Identification of Nrl1 Domains Responsible for Interactions with RNA-Processing Factors and Regulation of Nrl1 Function by Phosphorylation." International Journal of Molecular Sciences 22, no. 13 (June 29, 2021): 7011. http://dx.doi.org/10.3390/ijms22137011.

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Pre-mRNA splicing is a key process in the regulation of gene expression. In the fission yeast Schizosaccharomyces pombe, Nrl1 regulates splicing and expression of several genes and non-coding RNAs, and also suppresses the accumulation of R-loops. Here, we report analysis of interactions between Nrl1 and selected RNA-processing proteins and regulation of Nrl1 function by phosphorylation. Bacterial two-hybrid system (BACTH) assays revealed that the N-terminal region of Nrl1 is important for the interaction with ATP-dependent RNA helicase Mtl1 while the C-terminal region of Nrl1 is important for interactions with spliceosome components Ctr1, Ntr2, and Syf3. Consistent with this result, tandem affinity purification showed that Mtl1, but not Ctr1, Ntr2, or Syf3, co-purifies with the N-terminal region of Nrl1. Interestingly, mass-spectrometry analysis revealed that in addition to previously identified phosphorylation sites, Nrl1 is also phosphorylated on serines 86 and 112, and that Nrl1-TAP co-purifies with Cka1, the catalytic subunit of casein kinase 2. In vitro assay showed that Cka1 can phosphorylate bacterially expressed Nrl1 fragments. An analysis of non-phosphorylatable nrl1 mutants revealed defects in gene expression and splicing consistent with the notion that phosphorylation is an important regulator of Nrl1 function. Taken together, our results provide insights into two mechanisms that are involved in the regulation of the spliceosome-associated factor Nrl1, namely domain-specific interactions between Nrl1 and RNA-processing proteins and post-translational modification of Nrl1 by phosphorylation.

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Kang, Ji Young, Minchan Gil, and Kyung Eun Kim. "Neuropilin1 Expression Acts as a Prognostic Marker in Stomach Adenocarcinoma by Predicting the Infiltration of Treg Cells and M2 Macrophages." Journal of Clinical Medicine 9, no. 5 (May 12, 2020): 1430. http://dx.doi.org/10.3390/jcm9051430.

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Neuropilin1 (NRP1) plays a critical role in tumor progression and immune responses. Although the roles of NRP1 in various tumors have been investigated, the clinical relevance of NRP1 expression in stomach adenocarcinoma (STAD) has not been studied. To investigate the use of NRP1 as a prognostic biomarker of STAD, we analyzed NRP1 mRNA expression and its correlation with patient survival and immune cell infiltration using various databases. NRP1 mRNA expression was significantly higher in STAD than normal tissues, and Kaplan-Meier survival analysis showed that NRP1 expression was significantly associated with poor prognosis in patients with STAD. To elucidate the related mechanism, we analyzed the correlation between NRP1 expression and immune cell infiltration level. In particular, the infiltration of immune-suppressive cells, such as regulatory T (Treg) cells and M2 macrophage, was significantly increased by NRP1 expression. In addition, the expression of interleukin (IL)-35, IL-10, and TGF-β1 was also positively correlated with NRP1 expression, resulting in the immune suppression. Collectively in this study, our integrated analysis using various clinical databases shows that the significant correlation between NRP1 expression and the infiltration of Treg cells and M2 macrophage explains poor prognosis mechanism in STAD, suggesting the clinical relevance of NRP1 expression as a prognostic biomarker for STAD patients.

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Llarena, Marta, María J. Llama, and Juan L. Serra. "Purification and properties of NrtC and NrtD, the ATP-binding subunits of the ABC nitrate/nitrite transporter of Phormidium laminosum." Biochimica et Biophysica Acta (BBA) - General Subjects 1760, no. 12 (December 2006): 1819–26. http://dx.doi.org/10.1016/j.bbagen.2006.08.006.

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Götze, Tilmann, Maria Clara Soto-Bernardini, Mingyue Zhang, Hendrik Mießner, Lisa Linhoff, Magdalena M. Brzózka, Viktorija Velanac, et al. "Hyperactivity is a Core Endophenotype of Elevated Neuregulin-1 Signaling in Embryonic Glutamatergic Networks." Schizophrenia Bulletin 47, no. 5 (April 19, 2021): 1409–20. http://dx.doi.org/10.1093/schbul/sbab027.

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Abstract The neuregulin 1 (NRG1) ErbB4 module is at the core of an “at risk” signaling pathway in schizophrenia. Several human studies suggest hyperstimulation of NRG1-ErbB4 signaling as a plausible pathomechanism; however, little is known about the significance of stage-, brain area-, or neural cell type-specific NRG1-ErbB4 hyperactivity for disease-relevant brain endophenotypes. To address these spatiotemporal aspects, we generated transgenic mice for Cre recombinase-mediated overexpression of cystein-rich domain (CRD) NRG1, the most prominent NRG1 isoform in the brain. A comparison of “brain-wide” vs cell type-specific CRD-NRG1 overexpressing mice revealed that pathogenic CRD-NRG1 signals for ventricular enlargement and neuroinflammation originate outside glutamatergic neurons and suggests a subcortical function of CRD-NRG1 in the control of body weight. Embryonic onset of CRD-NRG1 in glutamatergic cortical networks resulted in reduced inhibitory neurotransmission and locomotor hyperactivity. Our findings identify ventricular enlargement and locomotor hyperactivity, 2 main endophenotypes of schizophrenia, as specific consequences of spatiotemporally distinct expression profiles of hyperactivated CRD-NRG1 signaling.

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Herzog, Birger, Caroline Pellet-Many, Gary Britton, Basil Hartzoulakis, and Ian C. Zachary. "VEGF binding to NRP1 is essential for VEGF stimulation of endothelial cell migration, complex formation between NRP1 and VEGFR2, and signaling via FAK Tyr407 phosphorylation." Molecular Biology of the Cell 22, no. 15 (August 2011): 2766–76. http://dx.doi.org/10.1091/mbc.e09-12-1061.

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In endothelial cells, neuropilin-1 (NRP1) binds vascular endothelial growth factor (VEGF)-A and is thought to act as a coreceptor for kinase insert domain-containing receptor (KDR) by associating with KDR and enhancing VEGF signaling. Here we report mutations in the NRP1 b1 domain (Y297A and D320A), which result in complete loss of VEGF binding. Overexpression of Y297A and D320A NRP1 in human umbilical vein endothelial cells reduced high-affinity VEGF binding and migration toward a VEGF gradient, and markedly inhibited VEGF-induced angiogenesis in a coculture cell model. The Y297A NRP1 mutant also disrupted complexation between NRP1 and KDR and decreased VEGF-dependent phosphorylation of focal adhesion kinase at Tyr407, but had little effect on other signaling pathways. Y297A NRP1, however, heterodimerized with wild-type NRP1 and NRP2 indicating that nonbinding NRP1 mutants can act in a dominant-negative manner through formation of NRP1 dimers with reduced binding affinity for VEGF. These findings indicate that VEGF binding to NRP1 has specific effects on endothelial cell signaling and is important for endothelial cell migration and angiogenesis mediated via complex formation between NRP1 and KDR and increased signaling to focal adhesions. Identification of key residues essential for VEGF binding and biological functions provides the basis for a rational design of antagonists of VEGF binding to NRP1.

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Moore, Jeff D., Edward P. Acosta, Victoria A. Johnson, Roland Bassett, Joseph J. Eron, Margaret A. Fischl, Mary C. Long, Daniel R. Kuritzkes, and Jean-Pierre Sommadossi. "Intracellular Nucleoside Triphosphate Concentrations in HIV-Infected Patients on Dual Nucleoside Reverse Transcriptase Inhibitor Therapy." Antiviral Therapy 12, no. 6 (August 2007): 981–86. http://dx.doi.org/10.1177/135965350701200615.

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Background Intracellular nucleoside reverse transcriptase inhibitor triphosphate (NRTI-TP) concentrations are crucial in suppressing HIV replication. Little is known about how commonly used dual-NRTI regimens affect the intracellular levels of NRTI-TPs, the active form of these drugs. This study investigates the effect of dual-NRTI therapy in intracellular NRTI-TP levels. Methods NRTI and NRTI-TP concentrations were evaluated in HIV-infected patients receiving either lamivudine (3TC) and stavudine (d4T) or lamivudine with zidovudine (ZDV); NRTI and NRTI-TP concentrations were determined using a validated HPLC/MS/MS method. Plasma HIV-1 RNA levels were determined at baseline and monthly to examine the relationship between NRTI-TP concentrations and plasma HIV-1 RNA. Results Forty-one subjects completed the study. 3TC-TP significantly increased between day 1 and week 28 from 1.48 to 5.00 pmol/106 peripheral blood mononuclear cells (PBMC; P<0.0001). NRTI-TP concentrations for d4T and ZDV did not significantly increase, with values at week 28 of 0.011 and 0.02 pmol/106 PBMC, respectively. Mean NRTI-TP/plasma ratios were 3%, 0.007% and 0.05% for 3TC, d4T and ZDV, respectively. Linear relationships were observed between ZDV- and 3TC-TP and changes in plasma HIV-1 RNA. Conclusion Of the three drugs studied, only 3TC-TP levels increased significantly between day 1 and week 28. ZDV-TP and 3TC-TP levels were unaffected by dual-NRTI therapy relative to monotherapy, regardless of the combination (3TC-ZDV or 3TC-d4T). Intracellular levels of d4T-TP were similar to previous reports for dual-NRTI therapy; however, in the case of d4T, these values appear lower than those achieved with d4T monotherapy.

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Chen, Linyun, Mandy Kwong, Ronghua Lu, David Ginzinger, Candy Lee, Laura Leung, and Jefferson Y. Chan. "Nrf1 Is Critical for Redox Balance and Survival of Liver Cells during Development." Molecular and Cellular Biology 23, no. 13 (July 1, 2003): 4673–86. http://dx.doi.org/10.1128/mcb.23.13.4673-4686.2003.

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ABSTRACT The Nrf1 transcription factor belongs to the CNC subfamily of basic leucine zipper proteins. Knockout of Nrf1 is lethal in mouse embryos, but nothing is known about the cell types that absolutely require its function during development. We show by chimera analysis that Nrf1 is essential for the hepatocyte lineage. Mouse embryonic stem cells lacking Nrf1 developed normally and contributed to most tissues in adult chimeras where Nrf1 is normally expressed. Nrf1-deficient cells contributed to fetal, but not adult, liver cells. Loss of Nrf1 function resulted in liver cell apoptosis in late-gestation chimeric fetuses. Fetal livers from mutant embryos exhibited increased oxidative stress and impaired expression of antioxidant genes, and primary cultures of nrf1−/− fetal hepatocytes were sensitive to tert-butyl hydroperoxide-induced cell death, suggesting that impaired antioxidant defense may be responsible for the apoptosis observed in the livers of chimeric mice. In addition, cells deficient in Nrf1 were sensitized to the cytotoxic effects of tumor necrosis factor (TNF). Our results provide in vivo evidence demonstrating an essential role of Nrf1 in the survival of hepatocytes during development. Our results also suggest that Nrf1 may promote cell survival by maintaining redox balance and protecting embryonic hepatocytes from TNF-mediated apoptosis during development.

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Bian, Yun, Maoyun Sun, Marcy Silver, Kalon K. L. Ho, Mark A. Marchionni, Anthony O. Caggiano, James R. Stone, et al. "Neuregulin-1 attenuated doxorubicin-induced decrease in cardiac troponins." American Journal of Physiology-Heart and Circulatory Physiology 297, no. 6 (December 2009): H1974—H1983. http://dx.doi.org/10.1152/ajpheart.01010.2008.

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Neuregulin-1 (NRG1) is a potential therapeutic agent for the treatment of doxorubicin (Dox)-induced heart failure. NRG1, however, activates the erbB2 receptor, which is frequently overexpressed in breast cancers. It is, therefore, important to understand how NRG1, via erbB2, protects the heart against Dox cardiotoxicity. Here, we studied NRG1-erbB2 signaling in Dox-treated mice hearts and in isolated neonatal rat ventricular myocytes (NRVM). Male C57BL/6 mice were treated with recombinant NRG1 before and daily after a single dose of Dox. Cardiac function was determined by catheterization. Two-week survival was analyzed by the Kaplan-Meier method. Cardiac troponins [cardiac troponin I (cTnI) and cardiac troponin T (cTnT)] and phosphorylated Akt protein levels were determined in mice hearts and in NRVM by Western blot analysis. Activation of caspases and ubiquitinylation of troponins were determined in NRVM by caspase assay and immunoprecipitation. NRG1 significantly improved survival and cardiac function in Dox-treated mice. NRG1 reduced the decrease in cTnI, cTnT, and cardiac troponin C (cTnC) and maintained Akt phosphorylation in Dox-treated mice hearts. NRG1 reduced the decrease in cTnI and cTnT mRNA and proteins in Dox-treated NRVM. Inhibition of erbB2, phosphoinositide 3-kinase (PI3K), Akt, and mTOR blocked the protective effects of NRG1 on cTnI and cTnT in NRVM. NRG1 significantly reduced Dox-induced caspase activation, which degraded troponins, in NRVM. NRG1 reduced Dox-induced proteasome degradation of cTnI. NRG1 attenuates Dox-induced decrease in cardiac troponins by increasing transcription and translation and by inhibiting caspase activation and proteasome degradation of troponin proteins. NRG1 maintains cardiac troponins by the erbB2-PI3K pathway, which may lessen Dox-induced cardiac dysfunction.

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Luo, Xiaoyang, Marguerite Prior, Wanxia He, Xiangyou Hu, Xiaoying Tang, Weizhen Shen, Satya Yadav, et al. "Cleavage of Neuregulin-1 by BACE1 or ADAM10 Protein Produces Differential Effects on Myelination." Journal of Biological Chemistry 286, no. 27 (May 16, 2011): 23967–74. http://dx.doi.org/10.1074/jbc.m111.251538.

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Neuregulin-1 (Nrg1) is encoded by a single gene and exists in naturally secreted and transmembrane isoforms. Nrg1 exerts its signaling activity through interaction with its cognate ErbB receptors. Multiple membrane-anchored Nrg1 isoforms, present in six different membrane topologies, must be processed by a protease to initiate a signaling cascade. Here, we demonstrate that BACE1 and ADAM10 can process type I and III Nrg1 at two adjacent sites. Our cleavage site mapping experiments showed that the BACE1 cleavage site is located eight amino acids downstream of the ADAM10 cleavage site, and this order of cleavage is the opposite of amyloid precursor protein cleavage by these two enzymes. Cleavages were further confirmed via optimized electrophoresis. Cleavage of type I or III Nrg1 by ADAM10 and BACE1 released a signaling-capable N-terminal fragment (ntf), either Nrg1-ntfα or Nrg1-ntfβ, which could similarly activate an ErbB receptor as evidenced by increased phosphorylation of Akt and ERK, two downstream signaling molecules. Although both Nrg1-ntfα and Nrg1-ntfβ could initiate a common signaling cascade, inhibition or down-regulation of ADAM10 alone in a co-culture system did not affect normal myelination, whereas specific inhibition of BACE1 impaired normal myelination. Thus, processing of Nrg1 by BACE1 appears to be more critical for regulating myelination. Our results imply that a significant inhibition of BACE1 could potentially impair Nrg1 signaling activity in vivo.

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Ghez, David, Yves Lepelletier, Sophie Lambert, Jean-Marie Fourneau, Vincent Blot, Sébastien Janvier, Bertrand Arnulf, et al. "Neuropilin-1 Is Involved in Human T-Cell Lymphotropic Virus Type 1 Entry." Journal of Virology 80, no. 14 (July 15, 2006): 6844–54. http://dx.doi.org/10.1128/jvi.02719-05.

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ABSTRACT Human T-cell lymphotropic virus type 1 (HTLV-1) is transmitted through a viral synapse and enters target cells via interaction with the glucose transporter GLUT1. Here, we show that Neuropilin-1 (NRP1), the receptor for semaphorin-3A and VEGF-A165 and a member of the immune synapse, is also a physical and functional partner of HTLV-1 envelope (Env) proteins. HTLV-1 Env and NRP1 complexes are formed in cotransfected cells, and endogenous NRP1 contributes to the binding of HTLV-1 Env to target cells. NRP1 overexpression increases HTLV-1 Env-dependent syncytium formation. Moreover, overexpression of NRP1 increases both HTLV-1 and HTLV-2 Env-dependent infection, whereas down-regulation of endogenous NRP1 has the opposite effect. Finally, overexpressed GLUT1, NRP1, and Env form ternary complexes in transfected cells, and endogenous NRP1 and GLUT1 colocalize in membrane junctions formed between uninfected and HTLV-1-infected T cells. These data show that NRP1 is involved in HTLV-1 and HTLV-2 entry, suggesting that the HTLV receptor has a multicomponent nature.

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Raimondi, Claudio. "Neuropilin-1 enforces extracellular matrix signalling via ABL1 to promote angiogenesis." Biochemical Society Transactions 42, no. 5 (September 18, 2014): 1429–34. http://dx.doi.org/10.1042/bst20140141.

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Neuropilin-1 (NRP1), together with neuropilin-2, belongs to the neuropilin family. Neuropilins are transmembrane proteins essential for vascular and neural development and act as co-receptors for secreted signalling molecules of the class 3 semaphorin and vascular endothelial growth factor A (VEGF-A) families. NRP1 promotes VEGF-A signal in blood vascular endothelium and semaphorin signal in lymphatic endothelium, by forming complexes with its co-receptors. Mouse mutant studies established that NRP1 expression is essential during development because mice lacking NRP1 expression die embryonically and show severe neuronal and cardiovascular defects. Even though the contribution of NRP1 to vascular development has been mainly ascribed to its function as a VEGF-A receptor, recent evidence suggests that NRP1 contributes to angiogenesis through VEGF-independent mechanisms. In the present paper, we provide an overview of NRP1 functions in the vasculature and discuss current knowledge of NRP1-dependent signalling in the endothelium.

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