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Content archived on 2024-04-30

Regulation of initiation of dna replication biochemical investigations usinga novel human cell-free system

Objective



Research objectives and content
Initiation of DNA replication is a key step in the regulation of the cell division cycle. The commitment to enter S-phase occurs late in G1 phase at the so-called restriction point. Both events are control in part by Gl cyclins and their cyclin- dependent kinases. Recently a novel cell free system has been developed in the group of Prof. Ron Laskey in which semi-conservative initiation of DNA replication was shown in G, phase nuclei from HeLa cells when incubated with cytosolic and nuclear factors from S-phase HeLa cells. Exploiting this novel human system this study will investigate coupling of signal transduction pathways to the initiation of DNA replication.
Training content (objective, benefit and expected impact)
Knowing that cyclin A/Cdk2 and cyclin E/Cdk2 stimulate replication synergistically, I will investigate the function of cyclin D1/Cdk4 complex and its relation to the other G1 cyclin complexes by adding the recombinant purified proteins. Furthermore novel factors such as a already characterised inhibitory activity in G1 nuclear extracts will be isolated by biochemical fractionation. Finally a sequence of action of regulatory initiation proteins might be defined. Understanding coupling of DNA replication to the cell cycle will have impact on the research of the genetic changes that lead to cancer.

Fields of science (EuroSciVoc)

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Funding Scheme

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RGI - Research grants (individual fellowships)

Coordinator

University of Cambridge
EU contribution
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Address
Tennis Court Road
CB2 1QR Cambridge
United Kingdom

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Total cost

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Participants (1)

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