E-GEOD-43384 - Budding yeast ATM/ATR contribute to meiotic double-strand-break (DSB) homeostasis by down-regulating Rec114, an essential component of the DSB-machinery

Status
Released on 1 June 2013, last updated on 2 May 2014
Organism
Saccharomyces cerevisiae
Samples (12)
Array (1)
Protocols (5)
Description
In most organisms, meiotic recombination begins with programmed DNA double strand break (DSB) formation by Spo11. Here, we present evidence that Tel1/Mec1, the budding yeast ATM/ATR, regulate DSB formation by phosphorylating Rec114, an essential Spo11-accessory protein. Analyses of a non-phosphorylatable- or phosphomimetic- alleles of rec114 revealed that DSB-dependent phosphorylation of Rec114 limited its association with DSB-hotspots resulting in reduction in DSB formation. Also observed were the impact of Rec114 phosphorylation on its homolog synapsis-associated removal from chromosomes and NDT80-dependent turnover. Specifically, we found that the synapsis- and NDT80-dependent Rec114 downregulation occurred later in the rec114 mutant with a reduced Spo11-catalysis, but earlier in the other with an enhanced catalysis, strongly implicating the existence of a feedback mechanism coupling the extent of Spo11-catalysis to Rec114 activity. Taken together, these observations suggest that three different mechanisms of down regulating Rec114 contribute to meiotic DSB homeostasis, a feedback mechanism to maintain the number of meiotic DSBs at the developmentally programmed level. 6 genome wide ChIPchip sets: 3 for meiotic DSB formation (Spo11-ChIP) and 3 for protein-DNA association (Rec114-ChIP), each for wild type and two mutants during meiosis (corresponding to the main Figure 3, as well as to Figures S3, S4, S5).
Experiment type
ChIP-chip by tiling array 
Contacts
Franz Klein <franz.klein@univie.ac.at>, Anthony L. Johnsona, Jesus Carballo, Marco Geyomat, Maria-Elisabetta Serrentinoc, Rita Cha, Silvia Panizza, Valerie Borde
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-43384.idf.txt
Sample and data relationshipE-GEOD-43384.sdrf.txt
Raw data (2)E-GEOD-43384.raw.1.zip, E-GEOD-43384.raw.2.zip
Additional data (1)E-GEOD-43384.additional.1.zip
Array designA-GEOD-7250.adf.txt
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