Genetics, Vol. 157, 103-118, January 2001, Copyright © 2001

Requirement for Three Novel Protein Complexes in the Absence of the Sgs1 DNA Helicase in Saccharomyces cerevisiae

Janet R. Mullena, Vivek Kaliramana, Samer S. Ibrahima, and Steven J. Brilla
a Department of Molecular Biology and Biochemistry, Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, New Jersey 08855

Corresponding author: Steven J. Brill, Department of Molecular Biology and Biochemistry, Rutgers University, 679 Hoes Lane, CABM, Piscataway, NJ 08854., brill{at}mbcl.rutgers.edu (E-mail)

Communicating editor: L. S. SYMINGTON

The Saccharomyces cerevisiae Sgs1 protein is a member of the RecQ family of DNA helicases and is required for genome stability, but not cell viability. To identify proteins that function in the absence of Sgs1, a synthetic-lethal screen was performed. We obtained mutations in six complementation groups that we refer to as SLX genes. Most of the SLX genes encode uncharacterized open reading frames that are conserved in other species. None of these genes is required for viability and all SLX null mutations are synthetically lethal with mutations in TOP3, encoding the SGS1-interacting DNA topoisomerase. Analysis of the null mutants identified a pair of genes in each of three phenotypic classes. Mutations in MMS4 (SLX2) and SLX3 generate identical phenotypes, including weak UV and strong MMS hypersensitivity, complete loss of sporulation, and synthetic growth defects with mutations in TOP1. Mms4 and Slx3 proteins coimmunoprecipitate from cell extracts, suggesting that they function in a complex. Mutations in SLX5 and SLX8 generate hydroxyurea sensitivity, reduced sporulation efficiency, and a slow-growth phenotype characterized by heterogeneous colony morphology. The Slx5 and Slx8 proteins contain RING finger domains and coimmunoprecipitate from cell extracts. The SLX1 and SLX4 genes are required for viability in the presence of an sgs1 temperature-sensitive allele at the restrictive temperature and Slx1 and Slx4 proteins are similarly associated in cell extracts. We propose that the MMS4/SLX3, SLX5/8, and SLX1/4 gene pairs encode heterodimeric complexes and speculate that these complexes are required to resolve recombination intermediates that arise in response to DNA damage, during meiosis, and in the absence of SGS1/TOP3.





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Genes Dev.Home page
V. Kaliraman, J. R. Mullen, W. M. Fricke, S. A. Bastin-Shanower, and S. J. Brill
Functional overlap between Sgs1-Top3 and the Mms4-Mus81 endonuclease
Genes & Dev., October 15, 2001; 15(20): 2730 - 2740.
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Proc. Natl. Acad. Sci. USAHome page
G. W. Birrell, G. Giaever, A. M. Chu, R. W. Davis, and J. M. Brown
A genome-wide screen in Saccharomyces cerevisiae for genes affecting UV radiation sensitivity
PNAS, October 12, 2001; (2001) 231366398.
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Proc. Natl. Acad. Sci. USAHome page
R. J. Bennett and J. C. Wang
Association of yeast DNA topoisomerase III and Sgs1 DNA helicase: Studies of fusion proteins
PNAS, September 5, 2001; (2001) 201387098.
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Genome ResHome page
L. Aravind and E. V. Koonin
Prokaryotic Homologs of the Eukaryotic DNA-End-Binding Protein Ku, Novel Domains in the Ku Protein and Prediction of a Prokaryotic Double-Strand Break Repair System
Genome Res., August 1, 2001; 11(8): 1365 - 1374.
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J. Biol. Chem.Home page
W. M. Fricke, V. Kaliraman, and S. J. Brill
Mapping the DNA Topoisomerase III Binding Domain of the Sgs1 DNA Helicase
J. Biol. Chem., March 16, 2001; 276(12): 8848 - 8855.
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Proc. Natl. Acad. Sci. USAHome page
R. J. Bennett and J. C. Wang
Association of yeast DNA topoisomerase III and Sgs1 DNA helicase: Studies of fusion proteins
PNAS, September 25, 2001; 98(20): 11108 - 11113.
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Proc. Natl. Acad. Sci. USAHome page
G. W. Birrell, G. Giaever, A. M. Chu, R. W. Davis, and J. M. Brown
A genome-wide screen in Saccharomyces cerevisiae for genes affecting UV radiation sensitivity
PNAS, October 23, 2001; 98(22): 12608 - 12613.
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