About: DNA binding, stress-induced expression and transactivation activity of p73 protein isoforms     Goto   Sponge   NotDistinct   Permalink

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Description
  • Protein p73, a member of p53 family of transcription factors, shares remarkable homology with p53 protein and is able to transactivate many of p53-downstream genes involved in regulation of cell cycle and apoptosis. Just like by p53, several p73 isoforms were identified, which differ in their structure, however little is known about their DNA-binding properties, mutual interplay and cooperation, which might influence gene expression. This project is focused (ii) on in vitro studies of recognition of various DNA substrates (supercoiled, linear, chemically modified DNA) by selected p73 and p53 isoforms and (ii) on studies of DNA-binding and transactivation activities of both exogenous and endogenous p73 and p53 isoforms in different cell lines (with defined p53 status) exposed to different stress conditions. These investigations will be performed primarily using immunoprecipitation of the p73- or p53-DNA complexes and EMSA techniques, primer extension and PCR based methods, p53/p73 promoter-specific reporter assays and western blot analysis of p53/p73 isoform specific expression. (en)
  • Protein p73 patří mezi transkripční faktory rodiny p53, vykazuje vysokou homologii s proteinem p53 a je schopen aktivovat celou řadu genů, které se podílejí na regulaci buněčného cyklu a apoptózy. Podobně jako u proteinu p53 bylo identifikováno několik izoforem proteinu p73, které se liší strukturou, ale dosud chybí komplexnější data o jejich DNA-vazebných vlastnostech, vzájemné interakci a kooperaci, které mohou mít vliv na genovou expresi. Tento projekt je zaměřen (i) na in vitro studie rozpoznání různých DNA substrátů (nadšroubovicová, lineární, chemicky poškozená DNA) vybranými izoformami proteinů p73 a p53, a (ii) na DNA interakce a transaktivační aktivity exogenních a endogenních izoforem p73 a p53 ve vybraných buněčných liniích (s definovaným statutem p53) vystavených různým stresovým podmínkám. V těchto experimentech budou využity primárně metody imunoprecipitace p73 nebo p53-DNA komplexů, EMSA, prodlužování primerů pomocí DNA-polymeráz a PCR metody, p73/p53 promotor-specifické reportérové fůze a western blot analýzy exprese p73/p53 izoforem.
Title
  • DNA binding, stress-induced expression and transactivation activity of p73 protein isoforms (en)
  • Vazba na DNA, stresem indukovaná exprese a transaktivační aktivita izoforem proteinu p73
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  • GAP301/11/2076
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  • protein p73 isoforms tumor suppression DNA-protein interactions stress gene expression transcription (en)
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