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dc.contributor.advisorHupp, Ted
dc.contributor.advisorBall, Kathryn
dc.contributor.authorMaslon, Magdalena Maria
dc.date.accessioned2012-10-10T09:32:50Z
dc.date.available2012-10-10T09:32:50Z
dc.date.issued2012-06-30
dc.identifier.urihttp://hdl.handle.net/1842/6477
dc.description.abstractInactivation of p53 by mutation occurs in half of human tumours. The majority of these mutations affect the DNA-binding core domain and hence impair DNA binding and hinder transcription of p53 target genes. A wealth of data indicates that even cancers carrying wild type p53 protein, evolve mechanisms to render the p53 pathway inactive. Thus, inactivation of the p53 response, either by mutation or the alternative mechanisms, allows unpurturbed tumour growth. Recent work identified Anterior Gradient-2 (AGR2) as a protein overexpressed in wild type p53 expressing tumours and it was subsequently shown that AGR2 inhibits p53 pathway. In this study I confirmed that AGR2 protein inhibits p53 and AGR2 depletion potentiates p53-dependent DNA damage response. As there were no physiological signals known that regulate the AGR2-p53 axis, here I set out to identify pathways that activate or inhibit AGR2. I found that transforming growth factor β(TGF- β) triggers AGR2 protein reduction and this is concomitant with the stabilisation and increased activity of p53 protein. TGF-β halts AGR2 transcription in a SMAD4- dependent manner and triggers AGR2 protein degradation involving an ATM kinase. I found that SMAD nuclear interacting protein (SNIP1) mediated the ATMdependent AGR2 degradation. Interestingly, SNIP1 overexpression by itself promoted AGR2 protein degradation. I found that AGR2 protein degradation was proteasome independent and involed autophagy-lysosomal degradation pathway. As the mechanism of p53 inhibition by AGR2 is not known, I reasoned that identifying interactors of AGR2 may potentially further our understanding of the mechanism accounting for AGR2-mediated p53 inhibtion. I isolated the ATP binding protein Reptin in the yeast two-hybrid system and subsequently validated it as an AGR2 binding partner. Mutations of the two ATP binding motifs in Reptin resulted in altered oligomerization and thermostability of Reptin and affected the AGR2-Reptin complex stability. I also identified the Reptin docking site and it was mapped to a divergent octapeptide loop. I found that AGR2-Reptin complex coimmunoprecipitated with the p53 protein. Subsequently, I showed that Reptin protein can influence p53 activity, and depending on local concentration, either inhibit the transcription of p53-genes or chaperone its DNA binding activity. Interestingly, I found that Reptin formed a stable complex, independent of AGR2, with p53 R175H, p53 F270A, p53 S269D and p53 S269A, which has implication for the Reptin function in cancers bearing mutant form of p53 protein.en
dc.language.isoenen
dc.publisherThe University of Edinburghen
dc.relation.hasversionMaslon, M.M. and T.R. Hupp, Drug discovery and mutant p53. Trends Cell Biol. 20(9): p. 542-55.en
dc.relation.hasversionMagdalena M. Maslon, Roman Hrstka, Borek Vojtesek and Ted R. Hupp, 'A Divergent Substrate-Binding Loop within the Pro-oncogenic Protein Anterior Gradient-2 Forms a Docking Site for Reptin', J. Mol. Biol. (2010) 404, 418–438, doi:10.1016/j.jmb.2010.09.035en
dc.relation.hasversionHrstka, R., Nenutil, R., Fourtouna, I., Maslon, M. M., Naughton, C., Langdon, S. et al. (2010). The prometastatic protein anterior gradient-2 predicts poor prognosis in tamoxifen-treated breast cancers. Oncogene, 29, 4838–4847.en
dc.relation.hasversionFourtouna, I., Murray, E., Nicholson, J., Maslon, M. M., Pang, L., Dryden, D.&Hupp, T. R. (2009). The anterior gradient-2 pathway as a model for developing peptide-aptamer anti-cancer drug leads that stimulate p53 function. Curr. Chem. Biol. 3, 124–137.en
dc.subjectp53en
dc.subjectreptinen
dc.subjectAGR2en
dc.subjectcanceren
dc.subjectTGF-βen
dc.titleRegulation and function of AGR2 and p53 pathwaysen
dc.typeThesis or Dissertationen
dc.type.qualificationlevelDoctoralen
dc.type.qualificationnamePhD Doctor of Philosophyen


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