PXD036747

PXD036747 is an original dataset announced via ProteomeXchange.

Title	H3K79 methyltransferase Dot1p/DOT1L is phosphorylated by proline-directed MAP kinases Hog1p and p38/MAPK11
Description	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.
HostingRepository	PRIDE
AnnounceDate	2024-01-29
AnnouncementXML	Submission_2024-01-28_18:32:40.210.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ryan Separovich
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	phosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-09-14 14:27:32	ID requested
⏵ 1	2024-01-28 18:32:41	announced

Separovich RJ, Karakatsanis NM, Gao K, Fuh D, Hamey JJ, Wilkins MR, Proline-directed yeast and human MAP kinases phosphorylate the Dot1p/DOT1L histone H3K79 methyltransferase. FEBS J, ():(2024) [pubmed]

10.1111/febs.17062;

submitter keyword: yeast, signalling, post-translational regulation, histone methylation,epigenetics, chromatin

Marc R. Wilkins
contact affiliation	Systems Biology Initiative, School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia
contact email	m.wilkins@unsw.edu.au
lab head
Ryan Separovich
contact affiliation	University of New South Wales
contact email	r.separovich@unsw.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD036747
     1. Label: PRIDE project
     2. Name: H3K79 methyltransferase Dot1p/DOT1L is phosphorylated by proline-directed MAP kinases Hog1p and p38/MAPK11