Double Mutants of <i>Saccharomyces cerevisiae</i> with Alterations in Global Genome and Transcription-Coupled Repair

Verhage, Richard A.; Gool, Alain J. van; Groot, Nanda de; Hoeijmakers, Jan H.J.; Putte, Pieter van de; Brouwer, Jaap

doi:10.1128/mcb.16.2.496

Cited by 92 publications

(104 citation statements)

References 36 publications

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“…Since RNAPII stalled at damage may make the damage more accessible to repair enzymes than damage within a nucleosome, this may be considered a form of TRC that is independent of CSB. A mechanism such as this may also constitute the coupling factor (rad 26)-independent pathway of TRC identified in yeast (33). The data presented in this report suggest three indirect roles for CSB in TRC.…”

Section: Discussionsupporting

confidence: 50%

Cockayne syndrome group B protein enhances elongation by RNA polymerase II

Selby

Sancar²

1997

Proc. Natl. Acad. Sci. U.S.A.

282

178

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Cockayne syndrome (CS) is characterized by impaired physical and mental development. Two complementation groups, CSA and CSB, have been identified. Here we report that the CSB gene product enhances elongation by RNA polymerase II. CSB stimulated the rate of elongation on an undamaged template by a factor of about 3. A thyminethymine cyclobutane dimer located in the template strand is known to be a strong block to transcription. Addition of CSB to the blocked polymerase resulted in addition of one nucleotide to the nascent transcript. Finally, addition of transcription factor IIS is known to cause polymerase blocked at a thymine-thymine cyclobutane dimer to digest its nascent transcript, and CSB counteracted this transcript shortening action of transcription factor IIS. Thus a deficiency in transcription elongation may contribute to the CS phenotype.

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Section: Discussionsupporting

confidence: 50%

Cockayne syndrome group B protein enhances elongation by RNA polymerase II

Selby

Sancar²

1997

Proc. Natl. Acad. Sci. U.S.A.

282

178

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“…Using this system we have demonstrated that extracts made from rad7, rad16, and rad23 strains failed to efficiently repair AAAF-induced DNA damage. In contrast, an extract made from a rad26 deletion strain retained wild-type levels of in vitro excision repair, in keeping with the demonstrated role of RAD26, the homolog of the human Cockayne syndrome group B gene, in transcription-coupled repair (44,54). Since our yeast transcription extracts also permit efficient transcription by RNA polymerase II, it may be possible, by using a DNA substrate containing an RNA polymerase II promoter, to employ this system to study the role of the Rad26 protein in transcription-coupled repair.…”

Section: Discussionsupporting

confidence: 62%

“…2A shows that extracts made from the rad7 and rad16 deletion strains were completely defective for NER in our cell-free system while the rad23 deletion strain had a reduced ability to repair damaged DNA. In contrast to these results with rad7, rad16, and rad23 strains, an extract prepared from a rad26 deletion strain retained its ability to perform NER in vitro, an expected result given the evidence that the Rad26 protein functions exclusively in the subpathway of transcription-coupled repair (44,53). To confirm that the deficient in vitro NER in the rad7 and rad16 extracts was not due simply to poor extract preparation from these particular strains, in vitro complementation experiments were also performed.…”

Section: Nucleotide Excision Repair In Yeast Cellmentioning

confidence: 94%

“…The functions of the proteins encoded by these three RAD genes remain largely unknown, although RAD7 and RAD16 have been shown to be necessary for repair of the non-transcribed strand of transcribed genes and for repair of transcriptionally silent regions of the genome (44,51). Rad23 may promote the formation of a repair complex (52).…”

Section: Nucleotide Excision Repair In Yeast Cellmentioning

confidence: 99%

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Assessing the Requirements for Nucleotide Excision Repair Proteins of Saccharomyces cerevisiae in an in Vitro System

Wong

Maniar

et al. 1996

Journal of Biological Chemistry

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Nucleotide excision repair (NER) is the primary mechanism by which both Saccharomyces cerevisiae and human cells remove the DNA lesions caused by ultraviolet light and other mutagens. This complex process involves the coordinated actions of more than 20 polypeptides. To facilitate biochemical studies of NER in yeast, we have established a simple protocol for preparing whole cell extracts which perform NER in vitro. As expected, this assay of in vitro repair was dependent on the products of RAD genes such as RAD14, RAD4, and RAD2. Interestingly, it was also dependent upon proteins encoded by the RAD7, RAD16, and RAD23 genes whose precise roles in NER are uncertain, but not the RAD26 gene whose product is believed to participate in coupling NER to transcription. Replication protein A (RPA/ Rpa), known to be required for NER in human cell extracts, was also shown by antibody inhibition and immunodepletion experiments to be required for NER in our yeast cell extracts. Moreover, yeast cells with temperature-sensitive mutations in the RFA2 gene, which encodes the 34-kDa subunit of Rpa, had increased sensitivity to UV and yielded extracts defective in NER in vitro. These data indicate that Rpa is an essential component of the NER machinery in S. cerevisiae as it is in mammalian cells. Nucleotide excision repair (NER)1 is a versatile DNA repair strategy found ubiquitously in prokaryotes and eukaryotes. NER is capable of removing a broad spectrum of DNA lesions caused by physical and chemical mutagens (1-5). Failure to remove DNA lesions from the genome as a result of defective NER may lead to cancer-susceptibility, as exemplified by the hereditary human disease xeroderma pigmentosum (XP) (2-5). NER involves the concerted actions of several different enzymatic activities and can be arbitrarily divided into the following steps: damage recognition, incision and excision of the lesion and its flanking DNA, and repair DNA synthesis to fill in the resulting single-stranded gap. Many of the proteins encoded by XP genes and their evolutionarily conserved RAD gene homologs in Saccharomyces cerevisiae are now known to function in the early steps of excision repair, participating in the removal of DNA lesions prior to the repair DNA synthesis step. Some of the proteins involved in the repair DNA synthesis step of excision repair also function in the replication of cellular DNA and include proteins such as proliferating cell nuclear antigen (6 -8) and replication protein A (9 -14).Both yeast and mammalian replication protein A (Rpa/RPA) are trimeric complexes consisting of polypeptides of approximately 70, 34, and 14 kDa (15-19). RPA, a single-stranded DNA-binding protein, might function in the DNA synthesis step of NER as it does in cellular and viral DNA replication. Recent studies (11-13, 20 -22) have suggested, however, that RPA participates in both the early (damage recognition, incision, excision) steps as well as the late (repair synthesis) step of NER. The human RPA complex is now known to interact directly with XPA (20 -23...

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“…This residual difference might be attributed to Rhp26-independent TCR. Brouwer and co-workers report that there is a Rad26-independent TCR in S. cerevisiae at the short regions downstream of the promoters of active genes (34,35). The contribution of this Rad26-independent TCR was expressed as the difference in UV survival between a NERϪ strain and a rad16 rad26 double mutant.…”

Section: Discussionmentioning

confidence: 99%

Transcription Dependence and the Roles of Two Excision Repair Pathways for UV Damage in Fission Yeast Schizosaccharomyces pombe

Yasuhira

Morimyo

Yasui

1999

Journal of Biological Chemistry

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Fission yeasts Schizosaccharomyces pombe possess two types of excision repair systems for UV-induced DNA damage, nucleotide excision repair (NER) and UVdamaged DNA endonuclease (UVDE)-dependent excision repair (UVER). Despite its high efficiency in damage removal, UVER defects have less effect on UV survival than NER defects. To understand the differential roles of two pathways, we examined strand-specific damage removal at the myo2 and rpb2 loci. Although NER removes cyclobutane pyrimidine dimers from the transcribed strand more rapidly than from the nontranscribed strand, UVER repairs cyclobutane pyrimidine dimers equally on both strands and at a much higher rate than NER. The low rate of damage removal from the nontranscribed strand in the absence of UVER indicates inefficient global genome repair (GGR) in this organism and a possible function of UVER as an alternative to GGR. Disruption of rhp26, the S. pombe homolog of CSB/ RAD26, eliminated the strand bias of NER almost completely and resulted in a significant increase of UV sensitivity of cells in a uvde⌬ background. We suggest that the combination of transcription-coupled repair of NER and rapid UVER contributes to UV survival in growing S. pombe cells, which is accomplished by transcriptioncoupled repair and GGR in other organisms.

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Double Mutants of Saccharomyces cerevisiae with Alterations in Global Genome and Transcription-Coupled Repair

Cited by 92 publications

References 36 publications

Cockayne syndrome group B protein enhances elongation by RNA polymerase II

Cockayne syndrome group B protein enhances elongation by RNA polymerase II

Assessing the Requirements for Nucleotide Excision Repair Proteins of Saccharomyces cerevisiae in an in Vitro System

Transcription Dependence and the Roles of Two Excision Repair Pathways for UV Damage in Fission Yeast Schizosaccharomyces pombe

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