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NatF contributes to an evolutionary shift in protein N-terminal acetylation and is important for normal chromosome segregation

Petra Van Damme UGent, Kristine Hole, Ana Pimenta-Marques, Kenny Helsens UGent, Joël Vandekerckhove UGent, Rui G Martinho, Kris Gevaert UGent and Thomas Arnesen (2011) PLOS GENETICS. 7(7).
abstract
N-terminal acetylation (N-Ac) is a highly abundant eukaryotic protein modification. Proteomics revealed a significant increase in the occurrence of N-Ac from lower to higher eukaryotes, but evidence explaining the underlying molecular mechanism(s) is currently lacking. We first analysed protein N-termini and their acetylation degrees, suggesting that evolution of substrates is not a major cause for the evolutionary shift in N-Ac. Further, we investigated the presence of putative N-terminal acetyltransferases (NATs) in higher eukaryotes. The purified recombinant human and Drosophila homologues of a novel NAT candidate was subjected to in vitro peptide library acetylation assays. This provided evidence for its NAT activity targeting Met-Lys- and other Met-starting protein N-termini, and the enzyme was termed Naa60p and its activity NatF. Its in vivo activity was investigated by ectopically expressing human Naa60p in yeast followed by N-terminal COFRADIC analyses. hNaa60p acetylated distinct Met-starting yeast protein N-termini and increased general acetylation levels, thereby altering yeast in vivo acetylation patterns towards those of higher eukaryotes. Further, its activity in human cells was verified by overexpression and knockdown of hNAA60 followed by N-terminal COFRADIC. NatF's cellular impact was demonstrated in Drosophila cells where NAA60 knockdown induced chromosomal segregation defects. In summary, our study revealed a novel major protein modifier contributing to the evolution of N-Ac, redundancy among NATs, and an essential regulator of normal chromosome segregation. With the characterization of NatF, the co-translational N-Ac machinery appears complete since all the major substrate groups in eukaryotes are accounted for.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
YEAST, PROTEOMICS, IDENTIFICATION, IN-VITRO, AMINO-ACID, GTPASE ARL3P, SACCHAROMYCES-CEREVISIAE, METHIONINE AMINOPEPTIDASE, DIPEPTIDE COMPOSITION, ALPHA-ACETYLTRANSFERASE COMPLEX
journal title
PLOS GENETICS
PLoS Genet.
volume
7
issue
7
article_number
e1002169
pages
19 pages
Web of Science type
Article
Web of Science id
000293338600018
JCR category
GENETICS & HEREDITY
JCR impact factor
8.694 (2011)
JCR rank
11/155 (2011)
JCR quartile
1 (2011)
ISSN
1553-7390
DOI
10.1371/journal.pgen.1002169
project
Bioinformatics: from nucleotids to networks (N2N)
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1890716
handle
http://hdl.handle.net/1854/LU-1890716
date created
2011-08-19 11:25:36
date last changed
2013-02-27 09:10:30
@article{1890716,
  abstract     = {N-terminal acetylation (N-Ac) is a highly abundant eukaryotic protein modification. Proteomics revealed a significant increase in the occurrence of N-Ac from lower to higher eukaryotes, but evidence explaining the underlying molecular mechanism(s) is currently lacking. We first analysed protein N-termini and their acetylation degrees, suggesting that evolution of substrates is not a major cause for the evolutionary shift in N-Ac. Further, we investigated the presence of putative N-terminal acetyltransferases (NATs) in higher eukaryotes. The purified recombinant human and Drosophila homologues of a novel NAT candidate was subjected to in vitro peptide library acetylation assays. This provided evidence for its NAT activity targeting Met-Lys- and other Met-starting protein N-termini, and the enzyme was termed Naa60p and its activity NatF. Its in vivo activity was investigated by ectopically expressing human Naa60p in yeast followed by N-terminal COFRADIC analyses. hNaa60p acetylated distinct Met-starting yeast protein N-termini and increased general acetylation levels, thereby altering yeast in vivo acetylation patterns towards those of higher eukaryotes. Further, its activity in human cells was verified by overexpression and knockdown of hNAA60 followed by N-terminal COFRADIC. NatF's cellular impact was demonstrated in Drosophila cells where NAA60 knockdown induced chromosomal segregation defects. In summary, our study revealed a novel major protein modifier contributing to the evolution of N-Ac, redundancy among NATs, and an essential regulator of normal chromosome segregation. With the characterization of NatF, the co-translational N-Ac machinery appears complete since all the major substrate groups in eukaryotes are accounted for.},
  articleno    = {e1002169},
  author       = {Van Damme, Petra and Hole, Kristine and Pimenta-Marques, Ana and Helsens, Kenny and Vandekerckhove, Jo{\"e}l and Martinho, Rui G and Gevaert, Kris and Arnesen, Thomas},
  issn         = {1553-7390},
  journal      = {PLOS GENETICS},
  keyword      = {YEAST,PROTEOMICS,IDENTIFICATION,IN-VITRO,AMINO-ACID,GTPASE ARL3P,SACCHAROMYCES-CEREVISIAE,METHIONINE AMINOPEPTIDASE,DIPEPTIDE COMPOSITION,ALPHA-ACETYLTRANSFERASE COMPLEX},
  language     = {eng},
  number       = {7},
  pages        = {19},
  title        = {NatF contributes to an evolutionary shift in protein N-terminal acetylation and is important for normal chromosome segregation},
  url          = {http://dx.doi.org/10.1371/journal.pgen.1002169},
  volume       = {7},
  year         = {2011},
}

Chicago
Van Damme, Petra, Kristine Hole, Ana Pimenta-Marques, Kenny Helsens, Joël Vandekerckhove, Rui G Martinho, Kris Gevaert, and Thomas Arnesen. 2011. “NatF Contributes to an Evolutionary Shift in Protein N-terminal Acetylation and Is Important for Normal Chromosome Segregation.” Plos Genetics 7 (7).
APA
Van Damme, Petra, Hole, K., Pimenta-Marques, A., Helsens, K., Vandekerckhove, J., Martinho, R. G., Gevaert, K., et al. (2011). NatF contributes to an evolutionary shift in protein N-terminal acetylation and is important for normal chromosome segregation. PLOS GENETICS, 7(7).
Vancouver
1.
Van Damme P, Hole K, Pimenta-Marques A, Helsens K, Vandekerckhove J, Martinho RG, et al. NatF contributes to an evolutionary shift in protein N-terminal acetylation and is important for normal chromosome segregation. PLOS GENETICS. 2011;7(7).
MLA
Van Damme, Petra, Kristine Hole, Ana Pimenta-Marques, et al. “NatF Contributes to an Evolutionary Shift in Protein N-terminal Acetylation and Is Important for Normal Chromosome Segregation.” PLOS GENETICS 7.7 (2011): n. pag. Print.