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Histone H3 threonine 11 phosphorylation is catalyzed directly by the meiosis-specific kinase Mek1 and provides a molecular readout of Mek1 activity in vivo

UID: 10673

Author(s): Kniewel, Ryan*, Murakami, Hajime, Liu, Yan, Ito, Masaru, Ohta, Kunihiro, Hollingsworth, Nancy M., Keeney, Scott * MSK affiliated

Description
Summary from the GEO: "Saccharomyces cerevisiae Mek1 is a CHK2/Rad53-family kinase that regulates meiotic recombination and progression upon its activation in response to DNA double-strand breaks (DSBs). The full catalog of direct Mek1 phosphorylation targets remains unknown. Here, we show that phosphorylation of histone H3 on threonine 11 (H3 T11ph) is induced by meiotic DSBs in S. cerevisiae and Schizosaccharomyces pombe. Molecular genetic experiments in S. cerevisiae confirmed that Mek1 is required for H3 T11ph and revealed that phosphorylation is rapidly reversed when Mek1 kinase is no longer active. Reconstituting histone phosphorylation in vitro with recombinant proteins demonstrated that Mek1 directly catalyzes H3 T11 phosphorylation. Mutating H3 T11 to nonphosphorylatable residues conferred no detectable meiotic defects, indicating that H3 T11ph is dispensable for Mek1 functions in controlling recombination. However, H3 T11ph provides an excellent marker of ongoing Mek1 kinase activity in vivo. Anti-H3 T11ph chromatin immunoprecipitation followed by deep sequencing demonstrated that H3 T11ph was highly enriched at presumed sites of attachment of chromatin to chromosome axes, gave a more modest signal along chromatin loops, and was present at still lower levels immediately adjacent to DSB hotspots. These localization patterns closely tracked the distribution of Red1 and Hop1, axis proteins required for Mek1 activation. These findings provide insight into the spatial disposition of Mek1 kinase activity and the higher order organization of recombining meiotic chromosomes.

ChIP-seq analysis of Red1 on budding yeast meiotic chromosomes. Distribution of Red1 at 3h after meiotic induction"
Subject of Study
Subject(s)
Access via GEO

Raw and BEDGRAPH sequencing data.
Accession #: GSE103823

Access via SRA

Red1-FLAG ChIP at 3h after meiotic induction.
Accession #: SRP117603

Access via BioProject

Additional information about overall initiative.
Accession #: PRJNA407108

Access Restrictions
Free to All
Access Instructions
The NCBI Gene Expression Omnibus, SRA, and BioProject databases provide open access to these files.
Associated Publications
Data Type
Equipment Used
Illumina MiSeq
Dataset Format(s)
TAR, RAW, BEDGRAPH
Dataset Size
11.5 Mb (TAR of BEDGRAPH), 99.9Mb (SRA)
Data Catalog Record Updated
2023-12-07