About the Project
During the cell division cycle, cells must accurately propagate both genetic information (chromosomal DNA sequence) and epigenetic information (DNA methylation, histone modification and higher-order chromatin structure). My lab has been using a proteomics-based approach to identify key proteins and protein complexes required for maintaining the integrity of genomic DNA through the processes of DNA replication. A key event is the licensing of replication origins, which occurs in late mitosis and G1 and results in the loading of Mcm2-7 onto replication origins to prepare them for replication in the upcoming S phase. The presence of Mcm2-7 on chromatin provides a platform for the assembly of other protein complexes onto DNA which coordinate various processes required for the accurate duplication of the genome. The project will involve the use of mass spectrometry to characterise several newly identified protein complexes that are assembled onto Mcm2-7 at replication origins. Basic biochemical analysis of the protein complexes will then be performed to determine how these protein complexes co-ordinate DNA replication and other cell cycle events. The project will use various model systems including the Xenopus cell-free system which supports efficient genomic DNA replication under normal cell cycle control.
References
Gillespie, P.J., Khoudoli, G.A., Stewart, G., Swedlow, J.R., and Blow, J.J. (2007). ELYS/MEL-28 chromatin association coordinates nuclear pore complex assembly and replication licensing. Curr Biol 17, 1657-1662. Khoudoli, G.A., Gillespie, P.J., Stewart, G., Andersen, J.S., Swedlow, J.R., and Blow, J.J. (2008). Temporal profiling of the chromatin proteome reveals system-wide responses to replication inhibition. Curr Biol 18, 838-843. Gambus, A., Khoudoli, G.A., Jones, R.C., and Blow, J.J. (2011). MCM2-7 form double hexamers at licensed origins in Xenopus egg extract. J Biol Chem 286, 11855-11864.
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