Investigating how post translational modifications of p97 affect its function and regulation during DNA synthesis

Davenport, Eve (2025). Investigating how post translational modifications of p97 affect its function and regulation during DNA synthesis. University of Birmingham. M.Res.

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Abstract

p97 is a AAA+ ATPase with very diverse functions, from DNA replication to proteolysis. However, it is unknown how p97 is regulated throughout the cell cycle to affect the variety of its substrates differently in the right place and at the right time. In DNA replication, it is involved in removing the helicase from the chromatin when replication has ended. A previous member in the lab, discovered that p97 has different post translational modifications in S and M phases of the cell cycle including phosphorylation and ubiquitination. His-tagged mutant p97 proteins were made that were either catalytically dead, phospho-dead, phospho-mimicking or ubiquitin dead. These were then tested in the cell free Xenopus egg extract to see how they affect the protein dynamics of the replication machinery components: Mcm7, PCNA, Psf2 and Cdc45 during DNA replication via chromatin isolations. The interactions of these mutants with some of p97’s cofactors were also studied via His-pulldowns and it was found that in S-phase, phosphorylation could increase interaction of p97 with Npl4 and p47, while in M-phase, it may also increase with p47 and Ufd1. When p97 is catalytically dead, it may increase interaction with Ufd1. Alongside this, RPA and WRN were identified as proteins that may interact with p97 during DNA replication.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

Supervisor(s):