1 / 21

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.

lali
Download Presentation

GSK3B and congenital heart defects

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  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

  21. Questions???

More Related