E-GEOD-46841 - DNA replication and DSB factor binding in the meiotic S-phase checkpoint in budding yeast

Status
Released on 20 September 2013, last updated on 3 June 2014
Organism
Saccharomyces cerevisiae
Samples (62)
Array (1)
Protocols (10)
Description
During gamete formation, crossover recombination must occur on replicated DNA to ensure proper chromosome segregation in the first meiotic division. We identified a Mec1/ATR-dependent replication checkpoint in budding yeast that prevented the earliest stage of recombination, the programmed induction of DNA double-strand breaks (DSBs), when pre-meiotic DNA replication was delayed. The checkpoint suppressed DSBs through three complementary mechanisms: inhibition of Mer2 phosphorylation by Dbf4-dependent Cdc7 kinase, preclusion of chromosomal loading of Rec114 and Mre11, and lowered abundance of the Spo11 nuclease. Without this checkpoint, cells formed DSBs on partially replicated chromosomes. Importantly, such DSBs frequently failed to be repaired and impeded further DNA synthesis, leading to a rapid loss in cell viability. We conclude that a checkpoint-dependent constraint of DSB formation to duplicated DNA is critical not only for meiotic chromosome assortment, but also to protect genome integrity during gametogenesis. DSB factor association was measured in wild-type and checkpoint mutants strains under non-inducing or replication checkpoint inducing conditions. Additionally, DNA replication and helicase loading were measured in a replication and checkpoint deficient strain (cdc6-mn).
Experiment types
comparative genomic hybridization by array, ChIP-chip by tiling array 
Contacts
Andreas Hochwagen, Hannah G Blitzblau
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-46841.idf.txt
Sample and data relationshipE-GEOD-46841.sdrf.txt
Raw data (2)E-GEOD-46841.raw.1.zip, E-GEOD-46841.raw.2.zip
Processed data (1)E-GEOD-46841.processed.1.zip
Array designA-GEOD-5991.adf.txt
Links