Molecular basis of cell cycle control: p300 and pRb

Chan, Ho Man (2000) Molecular basis of cell cycle control: p300 and pRb. PhD thesis, University of Glasgow.

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Printed Thesis Information: https://eleanor.lib.gla.ac.uk/record=b1942924

Abstract

The retinoblastoma protein (pRb) and p300 are transcriptional co-repressor and co-activator respectively, playing key roles in regulating the cell cycle. Both molecules are targeted by a number of viral oncoproteins, suggesting that the functional inactivation of these molecules is required for oncogenicity. Furthermore, p300 has an intrinsic histone acetyltransferase activity (HAT), which is crucial to its function. Here, studies were performed to understand how acetylation and de-acetylation may influence cell cycle regulation. I demonstrated that pRb is a substrate of p300 HAT activity. Furthermore, adenovirus E1A re-directs p300 HAT activity and enhances pRb acetylation. Acetylated pRb preferentially binds to proto-oncoprotein MDM2, indicating the functional relevance of this modification in regulating pRb function. Acetylation of pRb also influences its phosphorylation pattern, implying a certain level of 'cross-talk' between these two post-translational modification events. In an attempt to understand the underlying mechanism of p300-mediated transcriptional co-activation, I investigated its interaction with a newly identified binding-partner NAP-2 (nucleosome assembly protein 2). First, the domains of interaction between these two proteins were studied. Subsequently, both NAP-2 and p300 were found to augment in transcriptional regulation. Overall, the results suggested a mechanism by which p300 and NAP family proteins co-operate in chromatin-remodeling related mechanisms in regulating transcription. Finally, a series of pRb B pocket mutants were created, one of which was significantly compromised in binding to LXCXE peptide motif, including histone deacetylases (HDACs). pRb's growth suppressive function was correlated to its intrinsic repressive activity on transcription. At least in part, this intrinsic repressive activity was described by its association with HDACs. Using this mutant, I analysed the necessity of EQDAC association for pRb mediated G1 arrest. Surprisingly, the results suggested that HD AC may be dispensable for such function. Therefore, it readdresses the question of the functional significance of the pRb-HDAC interaction.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Keywords: Cellular biology, molecular biology.
Colleges/Schools: College of Medical Veterinary and Life Sciences
Supervisor's Name: La Thangue, Prof. Nicholas B.
Date of Award: 2000
Depositing User: Enlighten Team
Unique ID: glathesis:2000-72905
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 11 Jun 2019 11:06
Last Modified: 11 Aug 2021 10:33
URI: https://theses.gla.ac.uk/id/eprint/72905
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