Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

SETD3 protein is the actin-specific histidine N-methyltransferase

Sebastian Kwiatkowski, Agnieszka K. Seliga, Maria Veiga-da-Cunha, Didier Vertommen, Marianna Terreri, Takao Ishikawa, Iwona Grabowska, Adam K Jagielski, Jakub Drozak
doi: https://doi.org/10.1101/266882
Sebastian Kwiatkowski
1Department of Metabolic Regulation, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Agnieszka K. Seliga
1Department of Metabolic Regulation, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Maria Veiga-da-Cunha
2Walloon Excellence in Lifesciences and Biotechnology (WELBIO), Brussels, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Didier Vertommen
3Protein Phosphorylation Unit, de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marianna Terreri
1Department of Metabolic Regulation, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Takao Ishikawa
4Department of Molecular Biology and the University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Iwona Grabowska
5Department of Cytology, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Adam K Jagielski
1Department of Metabolic Regulation, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jakub Drozak
1Department of Metabolic Regulation, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

Abstract

Protein histidine methylation is rarely studied posttranslational modification of unknown biochemical importance. In vertebrates, only a few methylhistidne-containing proteins have been reported so far, including β-actin as an essential example. The evolutionary conserved methylation of β-actin H73 residue is catalyzed by a specific histidine N-methyltransferase that has never been identified molecularly. In the present investigation, we have purified actin-specific histidine N-methyltransferase from rat muscles about 1200-fold. Its activity was studied by the radiochemical assay employing either homogeneous recombinant human β-actin produced in E. coli or its mutated form exhibiting substitution of H73 by Ala residue (H73A) as substrates. Three polypeptides of ≈65, 75 and 90 kDa coeluting with the enzyme activity were identified in the preparation. Mass spectrometry analysis of these polypeptides resulted in the identification of SETD3 methyltransferase as the only plausible candidate. Rat SETD3 and its human ortholog were expressed in COS-7 cells, purified to homogeneity and shown to catalyze methylation of β-actin at H73 residue as confirmed by mass spectrometry analysis. The SETD3 enzyme was active towards a synthetic peptide corresponding to residues 69-77 of β-actin, but not to its mutated form exhibiting His-to-Ala substitution. Finally, Setd3-deficient HAP1 cells were devoid of methylated H73 in β-actin and exhibited phenotypic changes, including a decrease in F-actin content and an increased glycolytic activity. We conclude that SETD3 is the actin-specific histidine N-methyltransferase. The data show for the first time the molecular identity of protein histidine N-methyltransferase in vertebrates and throw new light on the substrate specificity of SET-domain-containing enzymes.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted February 20, 2018.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
SETD3 protein is the actin-specific histidine N-methyltransferase
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
SETD3 protein is the actin-specific histidine N-methyltransferase
Sebastian Kwiatkowski, Agnieszka K. Seliga, Maria Veiga-da-Cunha, Didier Vertommen, Marianna Terreri, Takao Ishikawa, Iwona Grabowska, Adam K Jagielski, Jakub Drozak
bioRxiv 266882; doi: https://doi.org/10.1101/266882
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
SETD3 protein is the actin-specific histidine N-methyltransferase
Sebastian Kwiatkowski, Agnieszka K. Seliga, Maria Veiga-da-Cunha, Didier Vertommen, Marianna Terreri, Takao Ishikawa, Iwona Grabowska, Adam K Jagielski, Jakub Drozak
bioRxiv 266882; doi: https://doi.org/10.1101/266882

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (3506)
  • Biochemistry (7348)
  • Bioengineering (5324)
  • Bioinformatics (20266)
  • Biophysics (10020)
  • Cancer Biology (7744)
  • Cell Biology (11306)
  • Clinical Trials (138)
  • Developmental Biology (6437)
  • Ecology (9954)
  • Epidemiology (2065)
  • Evolutionary Biology (13325)
  • Genetics (9361)
  • Genomics (12587)
  • Immunology (7702)
  • Microbiology (19027)
  • Molecular Biology (7444)
  • Neuroscience (41049)
  • Paleontology (300)
  • Pathology (1230)
  • Pharmacology and Toxicology (2138)
  • Physiology (3161)
  • Plant Biology (6861)
  • Scientific Communication and Education (1273)
  • Synthetic Biology (1897)
  • Systems Biology (5313)
  • Zoology (1089)