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PMID: 15601841 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Nse2, a component of the Smc5-6 complex, is a SUMO ligase required for the response to DNA damage.

Molecular and cellular biology ·Vol. 25 ·No. 1 ·2005-01-00 ·Pages 185-96

Andrews EA, Palecek J, Sergeant J, Taylor E, Lehmann AR, Watts FZ

Abstract

The Schizosaccharomyces pombe SMC proteins Rad18 (Smc6) and Spr18 (Smc5) exist in a high-M(r) complex which also contains the non-SMC proteins Nse1, Nse2, Nse3, and Rad62. The Smc5-6 complex, which is essential for viability, is required for several aspects of DNA metabolism, including recombinational repair and maintenance of the DNA damage checkpoint. We have characterized Nse2 and show here that it is a SUMO ligase. Smc6 (Rad18) and Nse3, but not Smc5 (Spr18) or Nse1, are sumoylated in vitro in an Nse2-dependent manner, and Nse2 is itself autosumoylated, predominantly on the C-terminal part of the protein. Mutations of C195 and H197 in the Nse2 RING-finger-like motif abolish Nse2-dependent sumoylation. nse2.SA mutant cells, in which nse2.C195S-H197A is integrated as the sole copy of nse2, are viable, whereas the deletion of nse2 is lethal. Smc6 (Rad18) is sumoylated in vivo: the sumoylation level is increased upon exposure to DNA damage and is drastically reduced in the nse2.SA strain. Since nse2.SA cells are sensitive to DNA-damaging agents and to exposure to hydroxyurea, this implicates the Nse2-dependent sumoylation activity in DNA damage responses but not in the essential function of the Smc5-6 complex.

MeSH Terms
Amino Acid Sequence Blotting, Western Cell Cycle Proteins/metabolism Cell Survival Chromosomal Proteins, Non-Histone/metabolism DNA Damage DNA Repair Dose-Response Relationship, Radiation Enzyme Inhibitors/pharmacology Gene Deletion Hydroxyurea/pharmacology Kinetics Molecular Sequence Data Mutation Plasmids/metabolism Protein Binding Protein Conformation Protein Structure, Tertiary Recombination, Genetic Schizosaccharomyces/enzymology,genetics Schizosaccharomyces pombe Proteins/metabolism Time Factors Ultraviolet Rays
Chemicals
Cell Cycle Proteins Chromosomal Proteins, Non-Histone Enzyme Inhibitors Nse2 protein, S pombe Schizosaccharomyces pombe Proteins Smc5 protein, S pombe smc6 protein, S pombe Hydroxyurea
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Andrews Emily A
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9RQ, United Kingdom.
Palecek Jan
Sergeant John
Taylor Elaine
Lehmann Alan R
Watts Felicity Z
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.1.185-196.2005
Published
2005-01-00
Pages
185-96
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC538766
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.25.1.185-196.2005 PMC Full Text View journal details
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