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Title [sv]
Strukturella och funktionella studier av DNA polymeras epsilon
Title [en]
Structural and functional studies of DNA polymerase epsilon
Abstract [sv]
The architecture of the eukaryotic replication fork is still not known. This project has previously contributed to today´s model suggesting that DNA polymerase epsilon (Pol-eps) participates in the synthesis of leading strand and DNA polymerase delta of the lagging strand. Genetic experiments have suggested that Mcm2-7 is the helicase which unwinds DNA at the replication fork, but in vitro biochemical experiments have only demonstrated a weak helicase activity. There may be proteins which modulate the helicase activity and we have purified the candidate proteins as well as all proteins likely needed for the leading strand synthesis. Our long-term objective is to understand how the replication fork functions on the molecular level, when efficiently replicating DNA with high fidelity. Our short-term objectives are to reconstitute the leading strand synthesis, obtain a high resolution structure of Pol-eps, explore if there could be an unidentified prostetic group on Pol-eps, define the biochemical and biological functions of the accessory subunits of Pol-eps. identify and study the effect of post-translational modifications on Pol-eps, carry out single turn-over experiments to study the chemistry by which Pol-eps build a new DNA strand. Leading strand synthesis in the presence of the helicase would allow novel questions to be asked, since all in vitro replication assays have been carried out with single-stranded DNA templates instead of natural double-stranded DNA templates.
Principal InvestigatorJohansson, Erik
Coordinating organisation
Umeå University
Funder
Period
2011-01-01 － 2013-12-31
Identifiers
DiVA, id: project:1090Project, id: 2010-05071_VR

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