PXD042599

PXD042599 is an original dataset announced via ProteomeXchange.

Title	Mechanisms of site-specific methylation by a highly conserved elongation factor 1A lysine methyltransferase
Description	Translation elongation factor 1A (eEF1A) is an essential and highly conserved protein required for protein synthesis in eukaryotes. In both Saccharomyces cerevisiae and human, five different methyltransferase enzymes methylate specific residues on eEF1A, making eEF1A the eukaryotic protein targeted by the highest number of dedicated methyltransferases. eEF1A methyltransferases are highly selective enzymes, only targeting eEF1A and each targeting just one or two specific residues in eEF1A. However, the mechanism of this selectivity remains poorly understood. Here we have used AlphaFold modelling in combination with crosslinking mass spectrometry (XL-MS) and enzyme mutagenesis to reveal how S. cerevisiae elongation factor methyltransferase 4 (Efm4) specifically methylates eEF1A at K316. We find that a unique beta-hairpin motif, which extends out from the core methyltransferase fold, is important for methylation of eEF1A K316 in vitro. An alanine mutation of a single residue on this beta-hairpin, F212, significantly reduces Efm4 activity in vitro and in yeast cells. We show that the equivalent residue in human eEF1A-KMT2 (METTL10), F220, is also important for its activity towards eEF1A in vitro. Lastly, we find that phosphorylation of eEF1A at S314 negatively crosstalks with Efm4-mediated methylation of K316. Our findings demonstrate how protein methyltransferases can be being highly selective towards a single residue on a single protein.
HostingRepository	PRIDE
AnnounceDate	2024-01-04
AnnouncementXML	Submission_2024-01-04_08:23:39.603.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Joshua Hamey
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monomethylated residue; phosphorylated residue
Instrument	Orbitrap Fusion Lumos; LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2023-05-31 20:35:44	ID requested
⏵ 1	2024-01-04 08:23:40	announced

Dataset with its publication pending

submitter keyword: Crosslinking mass spectrometry, Protein methyltransferase,Protein methylation

Marc Wilkins
contact affiliation	School of Biotechnology and Biomolecular Sciences, University of New South Wales, NSW, Australia
contact email	m.wilkins@unsw.edu.au
lab head
Joshua Hamey
contact affiliation	University of New South Wales
contact email	j.hamey@unsw.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042599
     1. Label: PRIDE project
     2. Name: Mechanisms of site-specific methylation by a highly conserved elongation factor 1A lysine methyltransferase