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GSK3B and congenital heart defects

GSK3B and congenital heart defects. Eva Dimitrova Image retrieved from : http://www.smashingapps.com/. Congenital heart defects introduction. 35,000 babies with congenital heart defects Li treatment for bipolar disorder contravercies Li and GSK3B.

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GSK3B and congenital heart defects

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  1. GSK3B and congenital heart defects Eva Dimitrova Image retrieved from : http://www.smashingapps.com/

  2. Congenital heart defects introduction • 35,000 babies with congenital heart defects • Li treatment for bipolar disorder contravercies • Li and GSK3B Bruneau, B. (February 2008). Review Article The developmental genetics of congenital heart disease. Nature. 451, 943-948

  3. GSK3B – serine/threonine kinase Glycogen SynthaseKinase 3 – S/T kinase Other substrates similar mechanism Two known isoforms GSK3α and GSK3β Key regulator of Wnt/beta-catenin pathway P GSK3B Inhibition GS GS P Image retrieved from: http://www.ebi.ac.uk

  4. GSK3B key regulator in Wnt/ beta-catenin pathway Wnt pathway in resting cells Wnt pathway in activated cells 1. Target processes • Differentiation • Proliferation • Cell migration • Cell polarity 2. Wnt /beta-catenin molecular mechanism • Resting cells– beta-catenin degradation • Activated cells – beta-catenin accumulation, gene transcription Image retrieved from: David M Roberts, Kevin C Slep & Mark Peifer Nature Structural & Molecular Biology 14, 463 - 465 (2007)

  5. GSK3B – homology and phylogeny Muscle Clustal W method Data created using http://www.phylogeny.fr/

  6. GSK3B mutations and related phenotypes Xenopus Xgsk3K/R (et al Pierce, S 1995) Mice Gsk3b-/- KO gene (et al Kerkela, R. 2008) Arabidopsis thaliana AtSK3-2 R178A (et al Claisse, G 2007) Drosophila – sgg-/- loss of function (et al Heslip,T.1997) Zebrafish GSK3-beta MO silencing

  7. Conserved sites in the serine/threonine kinase domain of GSK3B homologues Homo sapiens GSK3α Homo sapiens GSK3β Pan troglodytes GSK3-like protein (predicted) Canis familiaris GSK3-like protein (predicted) Danio rerio GSK3β Drosophila melanogaster Sgg Caenorhabditis elegans Gsk-3 Arabidopsis thaliana AtSK3-2 S/T Kinase domain ATP binding site and Active site Images retrieved from PROSITE: http://www.expasy.ch/prosite

  8. ATP-binding site and active site homology Homo sapiens GSK3α Homo sapiens GSK3β Pan troglodytes GSK3-like protein (predicted ) Canis familiaris GSK3-like protein (predicted) Danio rerio GSK3β Drosophila melanogaster Sgg Caenorhabditis elegans Gsk-3 Arabidopsis thaliana AtSK3-2 ATP-binding site and Active site Conserved distance between the two sites Images retrieved from PROSITE: http://www.expasy.ch/prosite

  9. Conclusion: players in Wnt pathway GSK3B interactions with other proteins Mus musculus Homo sapiens Data retrieved from :http://string.embl.de/

  10. Conclusion: players in Wnt pathway GSK3B interactions with other proteins Mus musculus Homo sapiens Data retrieved from :http://string.embl.de/

  11. GSK3B relationship to cardiac hypertrophy GSK3B - suggested to phosphorylate: 1. GATA4 2. NF-ATc 3. cMyc 4. Cyclin D1 Repression of cardiac hypertrophy Inhibition and nuclear exit Kerkela, R. (2008). Deletion of GSK-3 beta in mice leads to hypertrophic cardiomyopathy secondary to cardiomyoblast hyperproliferation. The journal of clinical investigation, 118, iss:11, 3609 -3618

  12. DNA Motifs on cDNA sequence Data retrieved from Motif search :http://motif.genome.jp/

  13. DNA Motifs on cDNA sequence Data retrieved from Motif search :http://motif.genome.jp/

  14. GATAs definition, structure, function GATA family – zinc finger transcription factors. They recognize the GATA motif in the promoter GATA4 -regulate genes involved in embryogenesis and in myocardial differentiation and function. Mutations in this gene have been associated with cardiac septal defects. GATA3 - Transcriptional activator which binds to the enhancer of the T-cell receptor alpha and delta genes. GATA2 - expressed in hematopoietic progenitors, promotes proliferation at the expense of differentiation GATA1 - erythroid development by regulating the switch of fetal hemoglobin to adult hemoglobin Data retrieved from GeneCards: http://www.genecards.org

  15. GSK3B interactions with GATA factors (Cyclin D1) Text mining Homology Databases Experiments Data retrieved from :http://string.embl.de/

  16. GSK3B interactions with GATA factors (Cyclin D1) Text mining Homology Databases Experiments Data retrieved from :http://string.embl.de/

  17. GATA protein motifs and homology Domains Zinc Finger DNA binding Pfam GATA-N transcription activation GATA1 GATA2 GATA3 GATA4 Data retrieved from SMART:http://smart.embl.de

  18. Microarray- GATA 1 knockdown and GSK3B expression Title: GDS1245 / 1437001_at / Gsk3b / Mus musculusSummary: Analysis of megakaryocytes lacking the transcription factor GATA-1. GATA-1 is required for the development of megakaryocytes and erythroid cells. Megakaryocytes obtained from 13.5 day C57BL/6 mutant embryos. Results provide insight into the role of GATA-1 in megakaryopoiesis. Data retrieved from : http://www.ncbi.nlm.nih.gov/geo/

  19. Future approaches Experiments for GSK3B and GATA4 interactions TAP-tag – specific protein interactions Y2H – no need to force nuclear entry - can even identify specific site interactions DNA-expression plasmid arrays GATA4 knock out or knock down experiments compare to GSK3B Model organisms: MO (morpholino) – early stages of development; easy to use RNAi – for early stages, knock out genes – zygotic expression

  20. References 1.Frame, S. (2001).GSK3 takes centre stage more than 20 years after its discovery.The Biochemical journal, 359, 1 -16.2. Kerkela, R. (2008). Deletion of GSK-3 beta in mice leads to hypertrophic cardiomyopathy secondary to cardiomyoblasthyperproliferation. The journal of clinical investigation, 118, iss:11, 3609 -36183. Science daily (2008, October 3). Loss of the protein target of lithium disrupts normal mouse embryonic heart development. Retrieved February 1, 2009 from http://www.sciencedaily.com4. Congenital heart defects. Retrieved January 31, 2009 from http://www.nhlbi.nih.gov/health/dci/index.html5. National center for biotechnology information http://www.ncbi.nlm.nih.gov/sites6. Brady, H. and Horgan, J.(1998, January). Lithium and the heart. Chest, 93.1,166-169.  7. Wnt/ beta-catenin signaling. Retrieved February 3, 2009 from http://www.cellsignal.com/reference/pathway/Wnt_beta_Catenin.html8.BLAST :http://blast.ncbi.nlm.nih.gov/Blast.cgi9. Homologene:http://www.ncbi.nlm.nih.gov/sites/entrez?db=homologene10 .Phylogeny fr: http://www.phylogeny.fr11. SMART: http://smart.embl-heidelberg.de12. Pfam: http://pfam.sanger.ac.uk13.Doble, B. & Woodgett, J. (2003). GSK-3: tricks of the trade for multi-tasking kinase. Journal of cell science 116, 1175-118614.Lee, H. & Tsai, J. (August, 3 2007). Glycogen synthasekinase 3α and 3β have distinct functioning during cardiogenesis of Zebrafish embryo. BMC developmental biology, 7:93.15. Claisse, G. & Charrier, B. (February, 15 2007). The Arabidopsis thaliana GSK3/Shaggy like kinase AtSK3-2 modulates floral cell expansion. Plant Mol Biol 64:113-124.16. Pierce, S. & Kimelman, D. (1995). Regulation of Spemann organizer formation by the intracellular kinase Xgsk-3. Development 121, 755-765.17. PROSITE: http://www.expasy.ch/prosite18. Bax,B.(December 2001).The structure of phosphorylated GSK-3 beta complexed with a peptide, FRATtide, that Inhibits beta-cateninphosphorylation. Structure.9, 1143-1152

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