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Mechanism of in vivo activation of the MutL-Exo1 complex for meiotic crossover formation

UID: 10610

Author(s): Sanchez, Aurore, Mu, Xiaojing*, Borde, Valerie, Keeney, Scott * MSK affiliated

Description
Summary from the GEO: "Programmed DNA double-strand breaks (DSBs) initiate meiotic recombination and their subsequent repair culminates in crossover (CO) formation. COs result from the asymmetric cleavage of double-Holliday junction (dHJ) intermediates, that requires the MutLγ endonuclease and a non-catalytic function of Exo1, an activity essential for fertility but at risk of generating unwanted chromosome rearrangements. Here we show how crossover formation by MutLγ is activated at the right time and at the right place. MutLγ forms a constitutive complex with Exo1, and in meiotic cells transiently contacts the upstream MutSγ (Msh4-Msh5) heterodimer. MutLγ associates with DSB hotspots only once recombination intermediates have been stabilized and engaged in the crossover repair pathway. MutLγ-Exo1 is recruited to DSB hotspots independently of the polo-like Cdc5 kinase, but to activate dHJ resolution, Cdc5 is recruited to the recombination intermediates and interacts individually with both MutLγ and Exo1, suggesting their direct modification. in vivo, MutLγ occupancy is restrained on recombination intermediates, and MutLγ associates with the vast majority of DSB hotspots, but at a lower frequency in centromeres, consistent with a strategy to reduce at-risk crossover events in these regions, and in late replicating regions. Our data highlight the tight temporal and spatial control of the activity of this constitutive, potentially harmful, nuclease."

Overall design from the GEO: "Two biological replicate Spo11-oligo maps of Saccharomyces cerevisiae strain mlh3 at meiotic 5 hr time point"
Subject of Study
Saccharomyces cerevisiae
Subject(s)
DNA Breaks, Double-Stranded
Meiosis
MutL Proteins
Protein Binding
Recombination, Genetic
Access via GEO

TAR of BigWig files
Accession #: GSE133108

Access via SRA

Two biological replicate Spo11-oligo maps of Saccharomyces cerevisiae
Accession #: SRP202027

Access via BioProject

Additional information about overall initiative.
Accession #: PRJNA549945

Access Restrictions
Free to All
Access Instructions
The NCBI Gene Expression Omnibus, SRA, and BioProject databases provide open access to these files.
Associated Publications
Sanchez A, Adam C, Rauh F, et al. Exo1 recruits Cdc5 polo kinase to MutLγ to ensure efficient meiotic crossover formation. Proceedings of the National Academy of Sciences of the United States of America. 2020; 117(48):30577-30588.
Data Type
Genomic
Equipment Used
Illumina HiSeq 2500

High throughput sequencing system

Software Used
SRA Toolkit

The NCBI SRA Toolkit enables reading ("dumping") of sequencing files from the SRA database and writing ("loading") files into the .sra format.

Dataset Format(s)
TAR, BIGWIG
Dataset Size
268.2 MB (TAR of BIGWIG), 1.64 Gb (SRA)
Data Catalog Record Updated
2023-12-07