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Interactions between dna-dependent protein kinase and components of the dna recombination/repair apparatus

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DNA-dependent protein Kinase (DNA-PK) plays a central role in two processes involving DNA double strand breaks (DSB): (i) repair of DSB induced by ionising radiation, and (ii) V(D)J recombination that occurs in developing of B and T lymphocytes. DNA-PK has two components: DNA-PK catalytic subunit (DNA-PKcs) and Ku antigen. DNA-PKcs only becomes active when the Ku component binds to DSBs.
Our poposal is to study the mechanism by which DNA-PK functions in these two processes. One attractive possibility is that DNA-PK, by protein-protein interactions, recruits the machinery necessary to carry out DSB repair or V(D)J recombination to the site of DNA damage. For this, I will try to identify in vivo interactions with human proteins using the "two hybrid" system. Hopefully, once identified the gene(s) that interacts with DNA-PK, I will map its genome position(s), and obtain antibodys against the protein(s) to study its cellular localization. Further in vitro studies will be done, by inmunoprecipitation, using antibodies against DNA-PK and co-purifing DNA-PK peptides.

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University of Cambridge
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