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Identifier uch.biology.msc//2001koutsodontis
Title Λειτουργικός χαρακτηρισμός των θέσεων πρόσδεσης του παράγοντα Sp1 στον υποκινητή του γονιδίου p21/WAF του ανθρώπου και μελέτη του μηχανισμού μεταγραφικής ενεργοποίησης από τις πρωτεΐνες SMAD και p300
Creator Koutsodontis, George
Abstract Διατμηματικό, συνεργαζόμενα Τμήματα Βιολογίας και Ιατρικής. Cell cycle progression is controlled by the cyclin-dependent kinases (CDKs). CDKS are the catalytic partners of the cyclins. p21 was identified as a CDK binding protein and proved to be a key factor in regulating cell cycle, as its expression following mitogenic stimuli results in G1 cell cycle arrest. The p21 gene is a target for diverse signals that induce growth arrest and differentiation. The transcriptional regulation of p21 is controlled by p53-dependent and p53-independent mechanisms. A variety of transcription factors bind to specific cis-acting elements located within the p21 promoter regulating the expression of the p21 gene in a p53-independent way. The region between -119/+1 of the p21 promoter contains six Sp1 binding sites and plays a major role in the regulation of p21 transcription. In the present work we constructed a series of point mutants of p21 promoter in each of Sp1 sites in order to study the importance of Sp1 sites in p21 regulation. Binding of Sp1 to the wild type and the mutated sites was confirmed by Gel Electrophoretic Mobility Shift Assay. Differences were observed in the activity of the mutated promoters using various cell lines, such as HepG2, COS-7 and HeLa. We further examined the role of the p300 factor and SMAD proteins for Sp1-mediated p21 transcription. Our results show that these sites are not of equal importance in p21 regulation and none of them is essential for Sp1-mediated transactivation. In addition all of the mutated promoter were transactivated by SMADs to the same extend (2 folds) suggesting that SMADs transactivate the p21 promoter by other factors that bind to the GC-rich elements of the proximal promoter Finally, there is evidence of a Sp1, p300 and SMAD multi-protein complex formation within the proximal region of p21 promoter, that controls the p21 transcription.
Issue date 2001 Ιατρκή-01-00
Date available 2000-11-07
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
Permanent Link https://elocus.lib.uoc.gr//dlib/d/0/7/metadata-dlib-2001koutsodontis.tkl Bookmark and Share
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