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Title: Modulation of MCM Helicase activity
Author: Satti, Rabab
ISNI:       0000 0004 2722 2957
Awarding Body: University of York
Current Institution: University of York
Date of Award: 2011
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The minichromosome maintenance (MCM) proteins are essential for the initiation and progression of DNA replication. MCMs are believed to function as the replicative helicase in archaea and eukaryotes. In the present studies, human MCM and archaeal MCM from Methanococcus maripaludis were purified from E.coli and assembled as complexes. The six MCMs from human eo-purified over multiples steps including three chromatographic steps when eo-expressed in E. coli suggesting that they form a heterohexamer complex. Reconstitution of in vitro helicase activity of the human complex and individual subunits of M maripaludis were described. Furthermore the effect of phosphorylation on human MCM complex and M maripaludis MCM A were also tested and the effect of phosphorylation was shown to be inhibitory on helicase activities. Different reaction conditions were monitored on MCM helicase activity and the most interesting was the effect of salts on helicase assays which showed that glutamate has a significant positive effect while chloride ions inhibit helicase activity. Putative kinases were purified from M maripaludis and Methanocaldococcus jannaschii and trials were conducted to assign them a role in regulating the DNA initiation process.
Supervisor: Chong, James Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available