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

Genetics - Tập 157 Số 1 - Trang 103-118 - 2001
Janet R. Mullen1, Vivek Kaliraman1, Samer Ibrahim1, Steven J. Brill1
1Department of Molecular Biology and Biochemistry, Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, New Jersey 08855

Tóm tắt

Abstract 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.

Từ khóa


Tài liệu tham khảo

Bailis, 1990, A defect in mismatch repair in Saccharomyces cerevisiae stimulates ectopic recombination between homeologous genes by an excision repair dependent process, Genetics, 126, 535, 10.1093/genetics/126.3.535

Bardwell, 1993, Yeast DNA recombination and repair proteins Rad1 and Rad10 constitute a complex in vivo mediated by localized hydrophobic domains, Mol. Microbiol., 8, 1177, 10.1111/j.1365-2958.1993.tb01662.x

Bardwell, 1994, Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease, Science, 265, 2082, 10.1126/science.8091230

Bender, 1991, Use of a screen for synthetic lethal and multicopy suppressor mutants to identify two new genes involved in morphogenesis in Saccharomyces cerevisiae, Mol. Cell. Biol., 11, 1295

Bennett, 1998, Purification and characterization of the Sgs1 DNA helicase activity of Saccharomyces cerevisiae, J. Biol. Chem., 273, 9644, 10.1074/jbc.273.16.9644

Bennett, 2000, Interaction between yeast Sgs1 helicase and DNA topoisomerase III, J. Biol. Chem., 275, 26898, 10.1016/S0021-9258(19)61459-6

Berg, 1996, The galvanization of biology: a growing appreciation for the roles of zinc, Science, 271, 1081, 10.1126/science.271.5252.1081

Boeke, 1987, 5-Fluoroorotic acid as a selective agent in yeast molecular genetics, Methods Enzymol., 154, 164, 10.1016/0076-6879(87)54076-9

Borden, 1996, The RING finger domain: a recent example of a sequence-structure family, Curr. Opin. Struct. Biol., 6, 395, 10.1016/S0959-440X(96)80060-1

Brill, 1988, Transcription-dependentDNA supercoiling in yeast DNA topoisomerase mutants, Cell, 54, 403, 10.1016/0092-8674(88)90203-6

Budd, 1995, DNA2 encodes a DNA helicase essential for replication of eukaryotic chromosomes, J. Biol. Chem., 270, 26766, 10.1074/jbc.270.45.26766

Chaganti, 1974, A manyfold increase in sister chromatid exchanges in Bloom’s syndrome lymphocytes, Proc. Natl. Acad. Sci. USA, 71, 4508, 10.1073/pnas.71.11.4508

Chakraverty, 1999, Defending genome integrity during DNA replication: a proposed role for RecQ family helicases, Bioessays, 21, 286, 10.1002/(SICI)1521-1878(199904)21:4<286::AID-BIES4>3.0.CO;2-Z

Courcelle, 1999, RecQ and RecJ process blocked replication forks prior to the resumption of replication in UV-irradiated Escherichia coli, Mol. Gen. Genet., 262, 543, 10.1007/s004380051116

Courcelle, 1997, recF and recR are required for the resumption of replication at DNA replication forks in Escherichia coli, Proc. Natl. Acad. Sci. USA, 94, 3714, 10.1073/pnas.94.8.3714

DiGate, 1988, Identification of a potent decatenating enzyme from Escherichia coli, J. Biol. Chem., 263, 13366, 10.1016/S0021-9258(18)37713-5

Ellis, 1995, The Bloom’s syndrome gene product is homologous to RecQ helicases, Cell, 83, 655, 10.1016/0092-8674(95)90105-1

Fujiwara, 1977, A retarded rate of DNA replication and normal level of DNA repair in Werner’s syndrome fibroblasts in culture, J. Cell Physiol., 92, 365, 10.1002/jcp.1040920305

Gangloff, 1994, The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase, Mol. Cell. Biol., 14, 8391

Gangloff, 1999, The essential role of yeast topoisomerase III in meiosis depends on recombination, EMBO J., 18, 1701, 10.1093/emboj/18.6.1701

Gangloff, 2000, Homologous recombination is responsible for cell death in the absence of the Sgs1 and Srs2 helicases, Nat. Genet., 25, 192, 10.1038/76055

Gebhart, 1988, Spontaneous and induced chromosomal instability in Werner syndrome, Hum. Genet., 80, 135, 10.1007/BF00702855

Gray, 1997, The Werner syndrome protein is a DNA helicase, Nat. Genet., 17, 100, 10.1038/ng0997-100

Guzder, 1994, RAD25 is a DNA helicase required for DNA repair and RNA polymerase II transcription, Nature, 369, 578, 10.1038/369578a0

Hand, 1975, A retarded rate of DNA chain growth in Bloom’s syndrome, Proc. Natl. Acad. Sci. USA, 72, 758, 10.1073/pnas.72.2.758

Harlow, 1988, Antibodies: A Laboratory Manual

Harmon, 1999, RecQ helicase and topoisomerase III comprise a novel DNA strand passage function: a conserved mechanism for control of DNA recombination, Mol. Cell, 3, 611, 10.1016/S1097-2765(00)80354-8

Johnson, 2000, Association of the Bloom syndrome protein with topoisomerase IIIα in somatic and meiotic cells, Cancer Res., 60, 1162

Karow, 1997, The Bloom’s syndrome gene product is a 3′-5′ DNA helicase, J. Biol. Chem., 272, 30611, 10.1074/jbc.272.49.30611

Karow, 2000, The Bloom’s syndrome gene product promotes branch migration of Holliday junctions, Proc. Natl. Acad. Sci. USA, 97, 6504, 10.1073/pnas.100448097

Kim, 1992, Identification of the yeast TOP3 gene product as a single strand-specific DNA topoisomerase, J. Biol. Chem., 267, 17178, 10.1016/S0021-9258(18)41910-2

Kitao, 1999, Mutations in RECQL4 cause a subset of cases of Rothmund-Thomson syndrome, Nat. Genet., 22, 82, 10.1038/8788

Lahaye, 1993, PIF1 DNA helicase from Saccharomyces cerevisiae. Biochemical characterization of the enzyme, J. Biol. Chem., 268, 26155, 10.1016/S0021-9258(19)74294-X

Lee, 1999, Requirement of yeast SGS1 and SRS2 genes for replication and transcription, Science, 286, 2339, 10.1126/science.286.5448.2339

Longtine, 1998, Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae, Yeast, 14, 953, 10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U

Lu, 1996, Human homologs of yeast DNA helicase, Nature, 383, 678, 10.1038/383678a0

Miozzo, 1998, Chromosomal instability in fibroblasts and mesenchymal tumors from two sibs with Rothmund-Thomson syndrome, Int. J. Cancer, 77, 504, 10.1002/(SICI)1097-0215(19980812)77:4<504::AID-IJC5>3.0.CO;2-Y

Mullen, 2000, Bipartite structure of the SGS1 DNA helicase from S. cerevisiae, Genetics, 154, 1101, 10.1093/genetics/154.3.1101

Murray, 1997, Role of Schizosaccharomyces pombe RecQ homolog, recombination, and checkpoint genes in UV damage tolerance, Mol. Cell. Biol., 17, 6868, 10.1128/MCB.17.12.6868

Park, 1992, RAD25 (SSL2), the yeast homolog of the human xeroderma pigmentosum group B DNA repair gene, is essential for viability, Proc. Natl. Acad. Sci. USA, 89, 11416, 10.1073/pnas.89.23.11416

Rodgers, 1996, A zinc-binding domain involved in the dimerization of RAG1, J. Mol. Biol., 260, 70, 10.1006/jmbi.1996.0382

Rong, 1993, Purification and characterization of the SRS2 DNA helicase of the yeast Saccharomyces cerevisiae, J. Biol. Chem., 268, 1252, 10.1016/S0021-9258(18)54067-9

Rong, 1991, The hyper-gene conversion hpr5-1 mutation of Saccharomyces cerevisiae is an allele of the SRS2/RADH gene, Genetics, 127, 75, 10.1093/genetics/127.1.75

Rose, 1990, Methods in Yeast Genetics

Rothstein, 1995, Hyper-recombination and Bloom’s syndrome: microbes again provide clues about cancer, Genome Res., 5, 421, 10.1101/gr.5.5.421

Salk, 1981, Cytogenetics of Werner’s syndrome cultured skin fibroblasts: variegated translocation mosaicism, Cytogenet. Cell Genet., 30, 92, 10.1159/000131596

Schiestl, 1988, RAD1, an excision repair gene of Saccharomyces cerevisiae, is also involved in recombination, Mol. Cell. Biol., 8, 3619

Schulz, 1994, The Saccharomyces PIF1 DNA helicase inhibits telomere elongation and de novo telomere formation, Cell, 76, 145, 10.1016/0092-8674(94)90179-1

Sikorski, 1989, A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae, Genetics, 12, 19, 10.1093/genetics/122.1.19

Sinclair, 1997, Extrachromosomal rDNA circles—a cause of aging in yeast, Cell, 91, 1033, 10.1016/S0092-8674(00)80493-6

Sinclair, 1997, Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants, Science, 277, 1313, 10.1126/science.277.5330.1313

Stewart, 1997, rqh1+, a fission yeast gene related to the Bloom’s and Werner’s syndrome genes, is required for reversible S phase arrest, EMBO J., 16, 2682, 10.1093/emboj/16.10.2682

Thomas, 1989, Elevated recombination rates in transcriptionally active DNA, Cell, 56, 619, 10.1016/0092-8674(89)90584-9

Umezu, 1993, RecQ DNA helicase of Escherichia coli, J. Mol. Biol., 230, 1145, 10.1006/jmbi.1993.1231

Umezu, 1990, Escherichia coli RecQ protein is a DNA helicase, Proc. Natl. Acad. Sci. USA, 87, 5363, 10.1073/pnas.87.14.5363

Vijayalaxmi, 1983, Bloom’s syndrome: evidence for an increased mutation frequency in vivo, Science, 221, 851, 10.1126/science.6879180

Wach, 1994, New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae, Yeast, 10, 1793, 10.1002/yea.320101310

Wang, 1991, DNA topoisomerases: Why so many?, J. Biol. Chem., 266, 6659, 10.1016/S0021-9258(20)89545-3

Watt, 1995, Sgs1: a eukaryotic homolog of E. coli RecQ that interacts with topoisom-erase II in vivo and is required for faithful chromosome segregation, Cell, 81, 253, 10.1016/0092-8674(95)90335-6

Watt, 1996, SGS1, a homologue of the Bloom’s and Werner’s syndrome genes, is required for maintenance of genome stability in Saccharomyces cerevisiae, Genetics, 144, 935, 10.1093/genetics/144.3.935

Wu, 2000, The Bloom’s syndrome gene product interacts with topoisomerase III, J. Biol. Chem., 275, 9636, 10.1074/jbc.275.13.9636

Xiao, 1998, Mms4, a putative transcriptional (co)activator, protects Saccharomyces cerevisiae cells from endogenous and environmental DNA damage, Mol. Gen. Genet., 257, 614, 10.1007/s004380050689

Yamagata, 1998, Bloom’s and Werner’s syndrome genes suppress hyperrecombination in yeast sgs1 mutant: implication for genomic instability in human diseases, Proc. Natl. Acad. Sci. USA, 95, 8733, 10.1073/pnas.95.15.8733

Yu, 1996, Positional cloning of the Werner’s syndrome gene, Science, 272, 258, 10.1126/science.272.5259.258

Thông tin
Thông tin xuất bản

Genetics

Tập 157 Số 1

103-118

Thông tin tác giả