Role of p65 cdc18 in cell cycle progression

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Project objective

The S. pombe p65cdcl8 is required for DNA replication and plays a major role in coordinating S phase and mitosis. The biochemical funcdon of the protein remains, however, unknown. The objective of the proposed research is to identify and characterise cdc18 interacting proteins, which will hopefully permit an insight into p65cdcl8 function. A biochemical approach will initially be followed: purification of p65cdcl8 from S. pombe extracts will allow the identification of associated factors and tbe cloning of the respective genes. The S. cerevisiae two-hybrid system will be used to test for direct association of the copurifying proteins with p65cdcl8 and to map the interaction domains. It can furthermore serve as an alternative approach to screen for p65cdcl8 interacting components. Characterisation of the putative interacting proteins identified will involve generation of null and conditional mutants and search for genetic interactions with cdc18 mutants. p65cdcl8 is likely to directly interact with the origin recognition complex. In vivo footprinting across S. pombe autonomously replicating sequences can be used to assess association of the identified proteins with origins of replication in cells.

Original text from CORDIS.

Participants

• Imperial Cancer Research Fund · LondonCoordinatorUnited Kingdom
• Not availableCity levelGreece

Links

• View on CORDIS