Dataset Information

H3K79 methyltransferase Dot1p/DOT1L is phosphorylated by proline-directed MAP kinases Hog1p and p38/MAPK11

ABSTRACT: Disruptor of telomeric silencing (Dot1p) is an exquisitely conserved histone methyltransferase, and is the sole enzyme responsible for H3K79 methylation in the budding yeast, Saccharomyces cerevisiae. It has been shown to be highly phosphorylated in vivo; however, the upstream kinases that act on Dot1p are unknown in yeast and all other eukaryotes. Here, we used in vitro and in vivo kinase discovery approaches to show that the mitogen-activated protein kinase Hog1p is a bona fide kinase of Dot1p methyltransferase. In in vitro kinase assays, Hog1p phosphorylates Dot1p at multiple sites, including at several proline-adjacent sites that are consistent with its known substrate preference. Hog1p kinase activity was specifically enhanced at these proline-adjacent sites on Dot1p upon its activation by the osmostress-responsive kinase, Pbs2p. We then show that genomic deletion of HOG1 markedly reduces phosphorylation at specific sites on Dot1p in vivo, thus providing evidence for Hog1p kinase activity on Dot1p in budding yeast cells. Phenotypic analysis of knockout and phosphosite mutant yeast strains revealed the importance of Hog1p-catalysed phosphorylation of Dot1p for cellular responses to ultraviolet-induced DNA damage. This kinase-substrate relationship is conserved in mammalian systems, where human DOT1L (ortholog of Dot1p) can be phosphorylated by the proline-directed kinase p38β/MAPK11 (ortholog of Hog1p) at multiple sites in vitro. Taken together, our findings establish Hog1p and p38β/MAPK11 as the first kinases known to act on H3K79 methyltransferase enzymes in any eukaryotic species.

INSTRUMENT(S): LTQ Orbitrap Velos, Q Exactive

ORGANISM(S): Homo Sapiens (human) Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Ryan Separovich

LAB HEAD: Marc R. Wilkins

PROVIDER: PXD036747 | Pride | 2024-01-29

REPOSITORIES: Pride

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Publications

Proline-directed yeast and human MAP kinases phosphorylate the Dot1p/DOT1L histone H3K79 methyltransferase.

Separovich Ryan J RJ Karakatsanis Nicola M NM Gao Kelley K Fuh David D Hamey Joshua J JJ Wilkins Marc R MR

The FEBS journal 20240125

Disruptor of telomeric silencing 1 (Dot1p) is an exquisitely conserved histone methyltransferase and is the sole enzyme responsible for H3K79 methylation in the budding yeast Saccharomyces cerevisiae. It has been shown to be highly phosphorylated in vivo; however, the upstream kinases that act on Dot1p are almost entirely unknown in yeast and all other eukaryotes. Here, we used in vitro and in vivo kinase discovery approaches to show that mitogen-activated protein kinase HOG1 (Hog1p) is a bona f ...[more]

PMID: 38270553

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Project description:In eukaryotes, regulation of mRNA translation enables a fast, localized and finely tuned expression of gene products. Within the translation process, the first stage of translation initiation is most rigorously modulated by the actions of eukaryotic initiation factors (eIFs) and their associated proteins. These 11 eIFs catalyze the joining of the tRNA, mRNA and rRNA into a functional translation complex. Their activity is influenced by a wide variety of extra- and intracellular signals, ranging from global, such as hormone signaling and unfolded proteins, to specific, such as single amino acid imbalance and iron deficiency. Their action is correspondingly comprehensive, in increasing or decreasing recruitment and translation of most cellular mRNAs, and specialized, in targeting translation of mRNAs with regulatory features such as a 5’ terminal oligopyrimidine tract (TOP), upstream open reading frames (uORFs), or an internal ribosomal entry site (IRES). In mammals, two major pathways are linked to targeted mRNA translation. The target of rapamycin (TOR) kinase induces translation of TOP and perhaps other subsets of mRNAs, whereas a family of eIF2 kinases does so with mRNAs containing uORFs or an IRES. TOR targets translation of mRNAs that code for proteins involved in translation, an action compatible with its widely accepted role in regulating cellular growth. The four members of the eIF2 kinase family increase translation of mRNAs coding for stress response proteins such as transcription factors and chaperones. Though all four kinases act on one main substrate, eIF2, published literature demonstrates both common and unique effects by each kinase in response to its specific activating stress. This suggests that the activated eIF2 kinases regulate the translation of both a global and a specific set of mRNAs. Up to now, few studies have attempted to test such a hypothesis; none has been done in mammals. We use array analysis to determine the global mRNA shift into polysomes following a stress response, and to compare the translational response following activation of GCN2 versus PERK, two of the four eIF2alpha kinases. This SuperSeries is composed of the following subset Series:; GSE11496: Expression data from Gcn2 wild-type and knockout mouse liver perfused with or without methionine; GSE11684: Expression data from Perk wild-type and knockout mouse liver perfused without or with 2,5-di-tert-butylhydroquinone Experiment Overall Design: Refer to individual Series

Dataset Information

H3K79 methyltransferase Dot1p/DOT1L is phosphorylated by proline-directed MAP kinases Hog1p and p38/MAPK11

Publications

Proline-directed yeast and human MAP kinases phosphorylate the Dot1p/DOT1L histone H3K79 methyltransferase.

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