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Genome-wide screen to identify novel factors in the replication checkpoint panthway : phd in molecular medicine

Jossen, Rachel <1978- >

Tesi o dissertazioni - 2009

1. ABSTRACT Chromosome integrity maintenance during DNA replication is essential for preventing genome rearrangements and cancer. When cells experience replication stress, the stability of stalled forks is controlled by the checkpoint response, which in S. cerevisiae requires Rad53 kinase. Hence Rad53 deficient cells are unable to recover from fork stalling induced by the replication inhibitor hydroxyurea. We designed a genome-wide screen aimed to identify pathways contributing to the viability of rad53 deficient cells experiencing replication stress. We generated an arrayed library containing all viable Knock Out strains carrying an inducible dominant negative allele (GALrad53-D339A) and screened for mutants able to grow in the presence of HU under inducing conditions. By this procedure we have identified 18 mutants, out of _ 4300 represented in the library, whose deletion rescues the checkpoint deficiency under replication stress. These suppressors could be grouped into two classes according to their ability to rescue the kinase deficient allele rad53-K227A [...]
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