Academic literature on the topic 'Human 5-aminolevulinate synthase 2 expression'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the lists of relevant articles, books, theses, conference reports, and other scholarly sources on the topic 'Human 5-aminolevulinate synthase 2 expression.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Journal articles on the topic "Human 5-aminolevulinate synthase 2 expression"
1
Salamin, Olivier, Emeric Gottardo, Céline Schobinger, Gemma Reverter-Branchat, Jordi Segura, Martial Saugy, Tiia Kuuranne, Jean-Daniel Tissot, Bernard Favrat, and Nicolas Leuenberger. "Detection of Stimulated Erythropoiesis by the RNA-Based 5'-Aminolevulinate Synthase 2 Biomarker in Dried Blood Spot Samples." Clinical Chemistry 65, no. 12 (December 1, 2019): 1563–71. http://dx.doi.org/10.1373/clinchem.2019.306829.
Full textAbstract:
Abstract BACKGROUND Despite implementation of the Athlete Biological Passport 10 years ago, blood doping remains difficult to detect. Thus, there is a need for new biomarkers to increase the sensitivity of the adaptive model. Transcriptomic biomarkers originating from immature reticulocytes may be reliable indicators of blood manipulations. Furthermore, the use of dried blood spots (DBSs) for antidoping purposes constitutes a complementary approach to venous blood collection. Here, we developed a method of quantifying the RNA-based 5′-aminolevulinate synthase 2 (ALAS2) biomarker in DBS. MATERIALS The technical, interindividual, and intraindividual variabilities of the method, and the effects of storage conditions on the production levels of ALAS2 RNA were assessed. The method was used to monitor erythropoiesis stimulated endogenously (blood withdrawal) or exogenously (injection of recombinant human erythropoietin). RESULTS When measured over a 7-week period, the intra- and interindividual variabilities of ALAS2 expression in DBS were 12.5%–42.4% and 49%, respectively. Following withdrawal of 1 unit of blood, the ALAS2 RNA in DBS increased significantly for up to 15 days. Variations in the expression level of this biomarker in DBS samples were more marked than those of the conventional hematological parameters, reticulocyte percentage and immature reticulocyte fraction. After exogenous stimulation of erythropoiesis via recombinant human erythropoietin injection, ALAS2 expression in DBS increased by a mean 8-fold. CONCLUSIONS Monitoring of transcriptomic biomarkers in DBS could complement the measurement of hematological parameters in the Athlete Biological Passport and aid the detection of blood manipulations.
2
FUDA, Hirotoshi, Chikara SHIMIZU, Young C. LEE, Harukuni AKITA, and Charles A. STROTT. "Characterization and expression of human bifunctional 3′-phosphoadenosine 5′-phosphosulphate synthase isoforms." Biochemical Journal 365, no. 2 (July 15, 2002): 497–504. http://dx.doi.org/10.1042/bj20020044.
Full textAbstract:
Sulphonation, a fundamental process essential for normal growth and development, requires the sulphonate donor molecule 3′-phosphoadenosine 5′-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the bifunctional enzyme PAPS synthase. In humans, two genes encode isoenzymes that are 77% identical at the amino acid level, and alternative splicing creates two subtypes of PAPS synthase 2. The question as to whether distinctions in amino acid composition are reflected in differences in activity has been examined. The specific activity of the PAPS synthase 2 subtypes is 10- to 15-fold higher than that for PAPS synthase 1. The greater catalytic efficiency of the PAPS synthase 2 subtypes is demonstrated further by the 3- to 6-fold higher kcat/Km ratios for ATP and inorganic sulphate as compared with the ratios for PAPS synthase 1. In humans, PAPS synthase 1 is expressed ubiquitously, and is the dominant isoform in most tissues, whereas expression of the PAPS synthase 2 subtypes is variable and tissue-specific. It is noteworthy that, similar to other human tissues, PAPS synthase 1 also appears to be the dominant isoform expressed in cartilage. The latter finding initially created a conundrum, since there is a specific human dwarfing disorder that is known to be caused by a mutation in the PAPS synthase 2 gene. This apparent enigma would seem to be resolved by examination of cartilage from guinea-pigs as an animal model. Similar to humans, cartilage from mature animals predominantly expresses PAPS synthase 1. In contrast, expression of PAPS synthase 1 is relatively low in the cartilage of immature guinea-pigs, including the growth plate of long bones, whereas PAPS synthase 2 is the highly expressed isoenzyme.
3
Moreau-Gaudry, Francois, Ping Xia, Gang Jiang, Natalya P. Perelman, Gerhard Bauer, James Ellis, Katherine H. Surinya, Fulvio Mavilio, Che-Kun Shen, and Punam Malik. "High-level erythroid-specific gene expression in primary human and murine hematopoietic cells with self-inactivating lentiviral vectors." Blood 98, no. 9 (November 1, 2001): 2664–72. http://dx.doi.org/10.1182/blood.v98.9.2664.
Full textAbstract:
AbstractUse of oncoretroviral vectors in gene therapy for hemoglobinopathies has been impeded by low titer vectors, genetic instability, and poor expression. Fifteen self- inactivating (SIN) lentiviral vectors using 4 erythroid promoters in combination with 4 erythroid enhancers with or without the woodchuck hepatitis virus postregulatory element (WPRE) were generated using the enhanced green fluorescent protein as a reporter gene. Vectors with high erythroid-specific expression in cell lines were tested in primary human CD34+ cells and in vivo in the murine bone marrow (BM) transplantation model. Vectors containing the ankyrin-1 promoter showed high-level expression and stable proviral transmission. Two vectors containing the ankyrin-1 promoter and 2 erythroid enhancers (HS-40 plus GATA-1 or HS-40 plus 5-aminolevulinate synthase intron 8 [I8] enhancers) and WPRE expressed at levels higher than the HS2/β-promoter vector in bulk unilineage erythroid cultures and individual erythroid blast-forming units derived from human BM CD34+ cells. Sca1+/lineage− Ly5.1 mouse hematopoietic cells, transduced with these 2 ankyrin-1 promoter vectors, were injected into lethally irradiated Ly5.2 recipients. Eleven weeks after transplantation, high-level expression was seen from both vectors in blood (63%-89% of red blood cells) and erythroid cells in BM (70%-86% engraftment), compared with negligible expression in myeloid and lymphoid lineages in blood, BM, spleen, and thymus (0%-4%). The I8/HS-40–containing vector encoding a hybrid human β/γ-globin gene led to 43% to 113% human γ-globin expression/copy of the mouse α-globin gene. Thus, modular use of erythroid-specific enhancers/promoters and WPRE in SIN-lentiviral vectors led to identification of high-titer, stably transmitted vectors with high-level erythroid-specific expression for gene therapy of red cell diseases.
4
Blouin, Jean-Marc, Cécile Ged, Magalie Lalanne, Isabelle Lamrissi-Garcia, Fanny Morice-Picard, Pierre Costet, Raêd Daher, et al. "Iron chelation rescues hemolytic anemia and skin photosensitivity in congenital erythropoietic porphyria." Blood 136, no. 21 (November 19, 2020): 2457–68. http://dx.doi.org/10.1182/blood.2020006037.
Full textAbstract:
Abstract Congenital erythropoietic porphyria (CEP) is an inborn error of heme synthesis resulting from uroporphyrinogen III synthase (UROS) deficiency and the accumulation of nonphysiological porphyrin isomer I metabolites. Clinical features are heterogeneous among patients with CEP but usually combine skin photosensitivity and chronic hemolytic anemia, the severity of which is related to porphyrin overload. Therapeutic options include symptomatic strategies only and are unsatisfactory. One promising approach to treating CEP is to reduce the erythroid production of porphyrins through substrate reduction therapy by inhibiting 5-aminolevulinate synthase 2 (ALAS2), the first and rate-limiting enzyme in the heme biosynthetic pathway. We efficiently reduced porphyrin accumulation after RNA interference–mediated downregulation of ALAS2 in human erythroid cellular models of CEP disease. Taking advantage of the physiological iron-dependent posttranscriptional regulation of ALAS2, we evaluated whether iron chelation with deferiprone could decrease ALAS2 expression and subsequent porphyrin production in vitro and in vivo in a CEP murine model. Treatment with deferiprone of UROS-deficient erythroid cell lines and peripheral blood CD34+-derived erythroid cultures from a patient with CEP inhibited iron-dependent protein ALAS2 and iron-responsive element–binding protein 2 expression and reduced porphyrin production. Furthermore, porphyrin accumulation progressively decreased in red blood cells and urine, and skin photosensitivity in CEP mice treated with deferiprone (1 or 3 mg/mL in drinking water) for 26 weeks was reversed. Hemolysis and iron overload improved upon iron chelation with full correction of anemia in CEP mice treated at the highest dose of deferiprone. Our findings highlight, in both mouse and human models, the therapeutic potential of iron restriction to modulate the phenotype in CEP.
5
Asano, Haruhiko, Xi Susan Li, and George Stamatoyannopoulos. "FKLF-2: a novel Krüppel-like transcriptional factor that activates globin and other erythroid lineage genes." Blood 95, no. 11 (June 1, 2000): 3578–84. http://dx.doi.org/10.1182/blood.v95.11.3578.011k48_3578_3584.
Full textAbstract:
FKLF-2, a novel Krüppel-type zinc finger protein, was cloned from murine yolk sac. The deduced polypeptide sequence of 289 amino acids has 3 contiguous zinc fingers at the near carboxyl-terminal end, an amino-terminal domain characterized by its high content of alanine and proline residues and a carboxyl-terminal domain rich in serine residues. By Northern blot hybridization, the human homologue of FKLF-2 is expressed in the bone marrow and striated muscles and not in 12 other human tissues analyzed. FKLF-2 is constitutively expressed in established cell lines with an erythroid phenotype, but it is inconsistently expressed in cell lines with myeloid or lymphoid phenotypes. The expression of FKLF-2 messenger RNA (mRNA) is up-regulated after induction of mouse erythroleukemia cells. In luciferase assays, FKLF-2 activates predominantly the γ, and to a lesser degree, the ɛ and β globin gene promoters. The activation of γ gene promoter does not depend on the presence of an HS2 enhancer. FKLF-2 activates the γ promoter predominantly by interacting with the γ CACCC box, and to a lesser degree through interaction with the TATA box or its surrounding DNA sequences. FKLF-2 also activated all the other erythroid specific promoters we tested (GATA-1, glycophorin B, ferrochelatase, porphobilinogen deaminase, and 5-aminolevulinate synthase). These results suggest that in addition to globin, FKLF-2 may be involved in activation of transcription of a wide range of genes in the cells of the erythroid lineage.
6
Asano, Haruhiko, Xi Susan Li, and George Stamatoyannopoulos. "FKLF-2: a novel Krüppel-like transcriptional factor that activates globin and other erythroid lineage genes." Blood 95, no. 11 (June 1, 2000): 3578–84. http://dx.doi.org/10.1182/blood.v95.11.3578.
Full textAbstract:
Abstract FKLF-2, a novel Krüppel-type zinc finger protein, was cloned from murine yolk sac. The deduced polypeptide sequence of 289 amino acids has 3 contiguous zinc fingers at the near carboxyl-terminal end, an amino-terminal domain characterized by its high content of alanine and proline residues and a carboxyl-terminal domain rich in serine residues. By Northern blot hybridization, the human homologue of FKLF-2 is expressed in the bone marrow and striated muscles and not in 12 other human tissues analyzed. FKLF-2 is constitutively expressed in established cell lines with an erythroid phenotype, but it is inconsistently expressed in cell lines with myeloid or lymphoid phenotypes. The expression of FKLF-2 messenger RNA (mRNA) is up-regulated after induction of mouse erythroleukemia cells. In luciferase assays, FKLF-2 activates predominantly the γ, and to a lesser degree, the ɛ and β globin gene promoters. The activation of γ gene promoter does not depend on the presence of an HS2 enhancer. FKLF-2 activates the γ promoter predominantly by interacting with the γ CACCC box, and to a lesser degree through interaction with the TATA box or its surrounding DNA sequences. FKLF-2 also activated all the other erythroid specific promoters we tested (GATA-1, glycophorin B, ferrochelatase, porphobilinogen deaminase, and 5-aminolevulinate synthase). These results suggest that in addition to globin, FKLF-2 may be involved in activation of transcription of a wide range of genes in the cells of the erythroid lineage.
7
Gianoukakis, Andrew G., H. James Cao, Timothy A. Jennings, and Terry J. Smith. "Prostaglandin endoperoxide H synthase expression in human thyroid epithelial cells." American Journal of Physiology-Cell Physiology 280, no. 3 (March 1, 2001): C701—C708. http://dx.doi.org/10.1152/ajpcell.2001.280.3.c701.
Full textAbstract:
KAT-50, an established human thyrocyte cell line, expresses constitutively high levels of prostaglandin endoperoxide H synthase-2 (PGHS-2), the inflammatory cyclooxygenase. Here, we examine primary human thyrocytes. We find that they, too, express PGHS-2 mRNA and protein under control culture conditions. A substantial fraction of the basal prostaglandin E2(PGE2) produced by these cells can be inhibited by SC-58125 (5 μM), a PGHS-2-selective inhibitor. Interleukin (IL)-1β (10 ng/ml) induces PGHS-2 expression and PGE2 production in primary thyrocytes. The induction of PGHS-2 and PGE2synthesis by IL-1β could be blocked by glucocorticoid treatment. Unlike KAT-50, most of the culture strains also express PGHS-1 protein. Our observations suggest that both cyclooxygenase isoforms may have functional roles in primary human thyroid epithelial cells, and PGHS-2 might predominate under basal and cytokine-activated culture conditions.
8
Noh, Seung-Jae, Y. Terry Lee, Colleen Byrnes, Antoinette Rabel, and Jeffery L. Miller. "Trafficking Kinesin Binding Protein Is Essential for Human Erythropoiesis." Blood 118, no. 21 (November 18, 2011): 683. http://dx.doi.org/10.1182/blood.v118.21.683.683.
Full textAbstract:
Abstract Abstract 683 Mitochondrial specialization in erythroblasts is important for efficient heme synthesis, with defects or reduced expression of several mitochondrial proteins causing anemia. Trafficking kinesin binding 2 (TRAK2) is known to participate in mitochondrial movement along microtubule by interacting with kinesin motor protein and making a complex with Miro that is localized on the mitochondrial outer membrane. Transcriptome data suggest that TRAK2 is highly and specifically expressed in early erythroid cells. Here the role of TRAK2 was studied among human CD34+ cells that were grown in ex vivo serum-free cultures supplemented with erythropoietin (EPO, total culture period 21 days). Quantitative PCR studies indicated that TRAK2 expression is highly regulated during erythropoiesis. Its expression pattern was nearly identical to aminolevulinate synthase 2, the erythroid specific enzyme for the committed step of the heme biosynthetic pathway, and mitoferrin 1, the erythroid specific mitochondrial iron transporter. Western analyses revealed that TRAK2 protein is detected as a doublet band with molecular weights of 130kD and 105kD. Mitochondrial co-localization of TRAK2 was verified by confocal microscopy in TRAK2-overexpressing K562 cells. To study a potential role of TRAK2 in erythropoiesis, TRAK2 expression was reduced in cultured human erythroid cells using lentiviral shRNA transduction. TRAK2 knockdown (TRAK2-KD) was confirmed by Western analysis in K562 cells. In primary erythroblasts, TRAK2-KD caused slight reduction of CD36+ immature erythroblasts at culture day 7 prior to the addition of EPO (CD36+ population 58% in control vs 40% in TRAK2-KD). After the addition of erythropoietin to the culture medium, TRAK2-KD severely restricted erythroblast proliferation (5.0 million cells/ml in control vs 0.25 million cells/ml in TRAK2-KD on culture day 18). Flow cytometric analyses showed that <1% of the CD36+ progenitors cells differentiated into glycophorin A erythroblasts compared with >90% in control cultures. Annexin-V staining indicated that more than 90% of cells had undergone apoptosis by day 14. These data suggest that TRAK2 expression is required for erythroid differentiation. As such, defects in TRAK2 expression should be considered in cases of unexplained anemia. The data also support the notion that mitochondrial location or mobility within erythroblasts may be important for iron trafficking or heme synthesis. Disclosures: No relevant conflicts of interest to declare.
9
Maharjan, Shreekrishna, Brahmaraju Mopidevi, Meenakshi Kaul Kaw, Nitin Puri, and Ashok Kumar. "Human aldosterone synthase gene polymorphism promotes miRNA binding and regulates gene expression." Physiological Genomics 46, no. 24 (December 15, 2014): 860–65. http://dx.doi.org/10.1152/physiolgenomics.00084.2014.
Full textAbstract:
Hypertension is a serious risk factor for myocardial infarction, heart failure, vascular disease, stroke, and renal failure. Like other complex diseases, hypertension is caused by a combination of genetic and environmental factors. The renin-angiotensin-aldosterone system plays an important role in the regulation of blood pressure. The octapeptide angiotensin II (ANG II) is one of the most active vasopressor agents and is obtained from the precursor molecule, angiotensinogen, by the combined proteolytic action of renin and angiotensin-converting enzyme. ANG II increases the expression of aldosterone synthase (coded by Cyp11B2 gene), which is the rate-limiting enzyme in the biosynthesis of aldosterone. Previous studies have shown that increased expression of aldosterone synthase increases blood pressure and cardiac hypertrophy in transgenic mice. Human Cyp11B2 gene has a T/C polymorphism at −344 positions in its 5′-untranslated region (UTR), and the −344T allele is associated with hypertension. Human Cyp11B2 gene also has an A/G polymorphism at 735 position in its 3′-UTR (rs28491316) that is in linkage disequilibrium with single nucleotide polymorphism at −344. We show here that 1) microRNA (miR)-766 binds to the 735G-allele and not the 735A-allele of the hCyp11B2 gene and 2) transfection of miR-766 reduces the human aldosterone synthase mRNA and protein level in human adrenocortical cells H295R. These studies suggest that miR-766 may downregulate the expression of human aldosterone synthase gene and reduce blood pressure in human subjects containing −344T allele.
10
M�ller-Decker, Karin, G�nther Reinerth, Peter Krieg, Regina Zimmermann, Helmut Heise, Christiane Bayerl, Friedrich Marks, and Gerhard F�rstenberger. "Prostaglandin-H-synthase isozyme expression in normal and neoplastic human skin." International Journal of Cancer 82, no. 5 (August 27, 1999): 648–56. http://dx.doi.org/10.1002/(sici)1097-0215(19990827)82:5<648::aid-ijc6>3.0.co;2-d.
Full textDissertations / Theses on the topic "Human 5-aminolevulinate synthase 2 expression"
1
Surinya, Katharina Helen. "Heme biosynthesis in erythroid cells : transcriptional egulation of the human 5=aminolevulinate synthase 2 gene /." Title page, contents and summary only, 1997. http://web4.library.adelaide.edu.au/theses/09PH/09phs961.pdf.
Full text
2
Dzikaitė, Vijolė. "Studies of proteins in heme and iron metabolism /." Stockholm, 2004. http://diss.kib.ki.se/2004/91-7349-762-2/.
Full text
3
Dell'Oso, Tania. "Regulation of human 5-aminolevulinate synthase 2 expression during erythroid differentiation and its role in x-linked sideroblastic anaemia." Thesis, 2003. http://hdl.handle.net/2440/92536.
Full textAbstract:
Haem is required for many cellular processes including haemoglobin synthesis in erythroid cells, where the majority of the haem in the body is produced and utilised. Its synthesis requires tight regulation to prevent toxic levels of free haem from arising. 5-Aminolevulinate synthase 2 (ALAS2) is the first and rate limiting enzyme in the biosynthesis of haem in erythroid cells. Thus, the regulation of ALAS2 expression is critical for maintaining and controlling haem production during the process of erythroid cell differentiation. A major aim of this study was to identify the regulatory elements within the ALAS2 gene that are involved in controlling transcription of ALAS2 during erythropoietin (Epo) stimulated erythroid differentiation. In order to investigate the regulation of ALAS2 transcription in the context of red blood cell maturation, an erythroid cell line, J2E, that terminally differentiates in response to Epo treatment was employed in this study. Human ALAS2 promoter deletion studies demonstrated that the first 293bp of the proximal promoter was sufficient to enhance transcription in response to Epo induced differentiation of the J2E cells. Introns 1 and 8 exhibited Epo responsive enhancer activity with intron 8 proving to be the stronger transcriptional activator in response to Epo. Transcription factor binding sites located in the 3' end of intron 8 that are critical to intron 8 Epo responsive enhancer activity were also identified. Preliminary studies on the effect of the coactivators CREB binding protein (CBP) and p300 on ALAS2 expression in response to Epo stimulation were conducted and suggested a potential involvement of these factors in regulating ALAS2 transcription. Defective haem synthesis, as a result of point mutations in the human ALAS2 gene, has been implicated in a blood disorder called X-linked sideroblastic anaemia (XLSA). XLSA is characterised by the presence of iron loaded mitochondria surrounding the nucleus in erythroblasts of the bone marrow. Anaemia, associated with a cycle of ineffective erythropoiesis that is linked to increased intestinal iron absorption and the secondary effect of iron overload is exhibited by XLSA patients. Point mutations in the ALAS2 gene of XLSA probands have been identified and two associated mutations, C1215G in exon 8 and Cl283T in exon 9 were selected as a basis for a murine model for XLSA. Thus, the aim of this project was to develop an animal model for XLSA to investigate the role of ALAS2 in this blood disorder and the associated defects in iron metabolism. A gene targeting approach using embryonic stem (ES) cells was employed and several strategies trialed. A potential ALAS2 targeted ES cell line containing the Cl159G point mutation in exon 8 (equivalent to the human mutation) was generated, with further characterisation required.Thesis (Ph.D.) -- University of Adelaide, Dept. of Molecular Biosciences, 2003
4
Surinya, Katharina Helen. "Heme biosynthesis in erythroid cells : transcriptional egulation of the human 5=aminolevulinate synthase 2 gene / a thesis submitted to the University of Adelaide for the degree of Doctor of Philosophy by Katharina Helen Surinya." Thesis, 1997. http://hdl.handle.net/2440/19094.
Full textAbstract:
Bibliography: leaves 193-234.xiv, 235, [147] leaves, [1] leaf of plates : ill. ; 30 cm.
Focusses on the elucidation of the regulatory mechanisms that control heme biosynthesis in eukaryotic cells.
Thesis (Ph.D.)--University of Adelaide, Dept. of Biochemistry, 1998?
Conference papers on the topic "Human 5-aminolevulinate synthase 2 expression"
1
Wilson, A. C., P. L. Kumar, M. M. Parker, I. Arora, J. D. Morrow, E. F. M. Wouters, R. Casaburi, et al. "5'-Aminolevulinate Synthase 2 (ALAS2) Expression Is a Potential Biomarker of Chronic Obstructive Pulmonary Disease (COPD) Cachexia." In American Thoracic Society 2019 International Conference, May 17-22, 2019 - Dallas, TX. American Thoracic Society, 2019. http://dx.doi.org/10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a5354.
Full textReports on the topic "Human 5-aminolevulinate synthase 2 expression"
1
Fridman, Eyal, and Eran Pichersky. Tomato Natural Insecticides: Elucidation of the Complex Pathway of Methylketone Biosynthesis. United States Department of Agriculture, December 2009. http://dx.doi.org/10.32747/2009.7696543.bard.
Full textAbstract:
Plant species synthesize a multitude of specialized compounds 10 help ward off pests. and these in turn may well serve as an alternative to synthetic pesticides to reduce environmental damage and health risks to humans. The general goal of this research was to perform a genetic and biochemical dissection of the natural-insecticides methylketone pathway that is specific to the glandular trichomes of the wild species of tomato, Solanumhabrochaites f. glabratum (accession PI126449). Previous study conducted by us have demonstrated that these compounds are synthesized de novo as a derivate pathway of the fatty acid biosynthesis, and that a key enzyme. designated MethylketoneSynthase 1 (MKS 1). catalyzes conversion of the intermediate B-ketoacyl- ACPs to the corresponding Cn-1 methylketones. The approach taken in this proposed project was to use an interspecific F2 population. derived from the cross between the cultivated lV182 and the wild species PIl26449. for three objectives: (i) Analyze the association between allelic status of candidate genes from the fatty acid biosynthesis pathway with the methylketone content in the leaves (ii) Perform bulk segregant analysis of genetic markers along the tomato genome for identifying genomic regions that harbor QTLs for 2TD content (iii) Apply differential gene expression analysis using the isolated glands of bulk segregant for identifying new genes that are involved in the pathway. The genetic mapping in the interspecific F2 population included app. 60 genetic markers, including the candidate genes from the FAS pathway and SSR markers spread evenly across the genome. This initial; screening identified 5 loci associated with MK content including the candidate genes MKS1, ACC and MaCoA:ACP trans. Interesting observation in this genetic analysis was the connection between shape and content of the glands, i.e. the globularity of the four cells, typical to the wild species. was associated with increased MK in the segregating population. In the next step of the research transcriptomic analysis of trichomes from high- and 10w-MK plants was conducted. This analysis identified a new gene, Methy1ketone synthase 2 (MKS2), whose protein product share sequence similarity to the thioesterase super family of hot-dog enzymes. Genetic analysis in the segregating population confirmed its association with MK content, as well as its overexpression in E. coli that led to formation of MK in the media. There are several conclusions drawn from this research project: (i) the genetic control of MK accumulation in the trichomes is composed of biochemical components in the FAS pathway and its vicinity (MKS 1 and MKS2). as well as genetic factors that mediate the morphology of these specialized cells. (ii) the biochemical pathway is now realized different from what was hypothesized before with MKS2 working upstream to I\1KS 1 and serves as the interface between primary (fatty acids) and secondary (MK) metabolism. We are currently testing the possible physical interactions between these two proteins in vitro after the genetic analysis showed clear epistatic interactions. (iii) the regulation of the pathway that lead to specialized metabolism in the wild species is largely mediated by transcription and one of the achievements of this project is that we were able to isolate and verify the specificity of the MKS1 promoter to the trichomes which allows manipulation of the pathways in these cells (currently in progress). The scientific implications of this research project is the advancement in our knowledge of hitherto unknown biochemical pathway in plants and new leads for studying a new family in plants (hot dog thioesterase). The agricultural and biotechnological implication are : (i) generation of new genetic markers that could assist in importing this pathway to cultivated tomato hence enhancing its natural resistance to insecticides, (ii) the discovery of MKS2 adds a new gene for genetic engineering of plants for making new fatty acid derived compounds. This could be assisted with the use of the isolated and verified MKS1 promoter. The results of this research were summarized to a manuscript that was published in Plant Physiology (cover paper). to a chapter in a proceeding book. and one patent was submitted in the US.