Journal
MOLECULAR AND CELLULAR BIOLOGY
Volume 25, Issue 15, Pages 6707-6721Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/MCB.25.15.6707-6721.2005
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- NIGMS NIH HHS [R01 GM047238] Funding Source: Medline
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Eukaryotes replicate DNA once and only once per cell cycle due to multiple, partially overlapping mechanisms efficiently preventing reinitiation. The consequences of reinitiation are unknown. Here we show that the induction of rereplication by mutations in components of the prereplicative complex (origin recognition complex [ORC], Cdc6, and minichromosome maintenance proteins) causes a cell cycle arrest with activated Rad53, a large-budded morphology, and an undivided nucleus. Combining a mutation disrupting the C1b5-Orc6 interaction (ORC6-rxl) and a mutation stabilizing Cdc6 (CDC6 Delta N7) causes a cell cycle delay with a similar phenotype, although this background is only partially compromised for rereplication control and does not exhibit overreplication detectable by fluorescence-activated cell sorting. We conducted a systematic screen that identified genetic requirements for the viability of these cells. ORC6-rxl CDC6 Delta NT cells depend heavily on genes required for the DNA damage response and for double-strand-break repair by homologous recombination. Our results implicate an Mre11-Mec1-dependent pathway in limiting the extent of rereplication.
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