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

Authors: Kniewel, R.; Murakami, H.; Liu, Y.; Ito, M.; Ohta, K.; Hollingsworth, N. M.; Keeney, S.
Article Title: Histone H3 threonine 11 phosphorylation is catalyzed directly by the meiosis-specific kinase Mek1 and provides a molecular readout of Mek1 activity in vivo
Abstract: 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confirmed that Mek1 is required for H3 T11ph and revealed that phosphorylation is rapidly reversed when Mek1 kinase is no longer active.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 defects in otherwise unperturbed meiosis, although the mutations modestly reduced spore viability in certain strains where Rad51 is used for strand exchange in place of Dmc1. H3 T11ph is therefore mostly dispensable for Mek1 function. 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. © 2017 by the Genetics Society of America.
Keywords: controlled study; nonhuman; chromosome; chromosome structure; meiosis; mitogen activated protein kinase kinase 1; in vivo study; in vitro study; enzyme activity; saccharomyces cerevisiae; chromatin; chromatin immunoprecipitation; histone h3; epitope; recombinant protein; western blotting; dna damage response; threonine; catalysis; single stranded dna; schizosaccharomyces pombe; gene cassette; histone methylation; histone phosphorylation; histone acetylation; priority journal; article; mek1
Journal Title: Genetics
Volume: 207
Issue: 4
ISSN: 0016-6731
Publisher: Genetics Society of America  
Date Published: 2017-12-01
Start Page: 1313
End Page: 1333
Language: English
DOI: 10.1534/genetics.117.300359
PROVIDER: scopus
PMCID: PMC5714450
PUBMED: 28986445
Notes: Article -- Export Date: 2 January 2018 -- Source: Scopus
Citation Impact
MSK Authors
  1. Scott N Keeney
    107 Keeney