Publication Type:

Journal Article

Source:

Molecular Biology of the Cell, Volume 19, Number 10, p.4099-4109 (2008)

URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-57349094259&partnerID=40&md5=8bdc2411054e3494bd677e56550e2c1d

Keywords:

Amino Acid, amino acid sequence, article, Carrier Proteins, Cell Cycle Proteins, cell viability, Chromosomal Proteins, chromosome segregation, dimerization, DNA damage, DNA repair, enzyme activity, Fungal Proteins, genomic instability, genotoxicity, heterodimer, Humans, immunoblotting, in vitro study, in vivo study, membrane protein, molecular cloning, Molecular Conformation, Molecular Sequence Data, Non-Histone, nonhuman, Nse1 RING like motif protein, Nuclear Proteins, nucleotide sequence, priority journal, protein domain, protein expression, protein function, protein motif, protein protein interaction, Protein Structure, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, sequence analysis, sequence homology, smc5 smc6 holocomplex, Stress, Tertiary, ubiquitin protein ligase E3, Ubiquitin-Protein Ligases, ubiquitination, unclassified drug

Abstract:

The Smc5-Smc6 holocomplex plays essential but largely enigmatic roles in chromosome segregation, and facilitates DNA repair. The Smc5-Smc6 complex contains six conserved non-SMC subunits. One of these, Nse1, contains a RING-like motif that often confers ubiquitin E3 ligase activity. We have functionally characterized the Nse1 RING-like motif, to determine its contribution to the chromosome segregation and DNA repair roles of Smc5-Smc6. Strikingly, whereas a full deletion of nse1 is lethal, the Nse1 RING-like motif is not essential for cellular viability. However, Nse1 RING mutant cells are hypersensitive to a broad spectrum of genotoxic stresses, indicating that the Nse1 RING motif promotes DNA repair functions of Smc5-Smc6. We tested the ability of both human and yeast Nse1 to mediate ubiquitin E3 ligase activity in vitro and found no detectable activity associated with full-length Nse1 or the isolated RING domains. Interestingly, however, the Nse1 RING-like domain is required for normal Nse1-Nse3-Nse4 trimer formation in vitro and for damage-induced recruitment of Nse4 and Smc5 to subnuclear foci in vivo. Thus, we propose that the Nse1 RING-like motif is a protein-protein interaction domain required for Smc5-Smc6 holocomplex integrity and recruitment to, or retention at, DNA lesions. © 2008 by The American Society for Cell Biology.

Notes:

cited By (since 1996)18

Cite this Research Publication

Sa Pebernard, Perry, J. J. Pab, Tainer, J. Aac, and Boddy, M. Na, “Nse1 RING-like domain supports functions of the Smc5-Smc6 holocomplex in genome stability”, Molecular Biology of the Cell, vol. 19, pp. 4099-4109, 2008.