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Content archived on 2024-04-30

Structure, stability, folding and function of the cancer susceptibility gene product brca 1

Objective



Research objectives and content
Soquence analysis of BRCA1, a cancer susceptibility gene responsible for tne majority of inherited breast cancers, lead to the discovery of a conserved domain, first described in the carboxyl terminus of BRCA1, a p53 binding protein (53BP1), and the yeast DNA repair protein RAD9. The BRCI domain (for BRCAl C-terminus) revealed a large family of domains that occur predominantly in proteins involved in cell cycle checkpoint functions responsive to DNA damage. The BRCI domain consists of 90 100 amino acid residues and occurs as a tandem repeat at the carboxy terminal of many proteins, but has been observed also as a single copy.
Determination of the three dimensional structure of the BRCT motif will be performed using nuclear magnetic resonance spectroscopy on the single BRCT domain of the telomere binding protein RAP1. In order to analyse the key residues that are important for specifying the native fold of the domain, protein engineering will be combined with stability measurements using fluorescence spectroscopy, circular dichroism and differential scanning calorimetry. A comparative study of the BRCT domain in different proteins will also be performed. The folding pathway will be characterized by fast kinetik techniques (stopped flow fluorescence). Functional studies will focus on binding studies with p53. This work should help unravel the functions of the BRCT domain and its role in the cell cycle and the evolution of cancer.
Training content (objective, benefit and expected impact)
The Cambridge Centre for Protdn Engineering is an Interdisciplinary Research Centre, directed by Aof A. R Fersht, FRS, which has considerable experience in the investigation of protein function, structure, stability and folding. It is therefore a privileged place to be trained in a wide variety of techniques by highly skilled individuals. The expertise of the Centre of Protein Engineering also favors contacts with other laboratories and research centres in Cambridge, in particular the Laboratory of Molecular Biology and the E.M.B.L. outstation. I hope, therefore, to benefit from this very stimulating environment and to acquire expertise in a range of techniques in order to obtain a PhD and continue my scientific education.

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Coordinator

MRC Centre for Protein Engineering
EU contribution
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Address
Hills Road
CB2 2QH Cambridge
United Kingdom

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