Requirement of Rad5 for DNA Polymerase ζ-Dependent Translesion Synthesis in <i>Saccharomyces cerevisiae</i>

Pagès, Vincent; Bresson, Anne; Acharya, Narottam; Prakash, Satya; Fuchs, Robert P. P.; Prakash, Louise

doi:10.1534/genetics.108.091066

Cited by 72 publications

(78 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Since little is known about the mechanistic details of errorfree tolerance, it is conceivable that Rev1 is capable of recruiting proteins involved in error-free tolerance in addition to the influence that polyubiquitinated PCNA has in signaling that pathway. This could be similar to the suspected structural role of Rad5 in Pol z-dependent TLS (Pages et al 2008). In contrast, the catalytic dead Rev1 does not seem to have a role in facilitating NER, because the catalytic dead rev1 strain is not much more sensitive in the rev1Drad14D background relative to the rev1D background ( Figure 5A vs. Figure 1D).…”

Section: Discussionsupporting

confidence: 53%

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Wiltrout¹,

Walker

2011

Genetics

View full text Add to dashboard Cite

A cell's ability to tolerate DNA damage is directly connected to the human development of diseases and cancer. To better understand the processes underlying mutagenesis, we studied the cell's reliance on the potentially error-prone translesion synthesis (TLS), and an error-free, template-switching pathway in Saccharomyces cerevisiae. The primary proteins mediating S. cerevisiae TLS are three DNA polymerases (Pols): Rev1, Pol z (Rev3/7), and Pol h (Rad30), all with human homologs. Rev1's noncatalytic role in recruiting other DNA polymerases is known to be important for TLS. However, the biological significance of Rev1's unusual conserved DNA polymerase activity, which inserts dC, is much less well understood. Here, we demonstrate that inactivating Rev1's DNA polymerase function sensitizes cells to both chronic and acute exposure to 4-nitroquinoline-1-oxide (4-NQO) but not to UV or cisplatin. Full Rev1-dependent resistance to 4-NQO, however, also requires the additional Rev1 functions. When error-free tolerance is disrupted through deletion of MMS2, Rev1's catalytic activity is more vital for 4-NQO resistance, possibly explaining why the biological significance of Rev1's catalytic activity has been elusive. In the presence or absence of Mms2-dependent error-free tolerance, the catalytic dead strain of Rev1 exhibits a lower 4-NQO-induced mutation frequency than wild type. Furthermore, Pol z, but not Pol h, also contributes to 4-NQO resistance. These results show that Rev1's catalytic activity is important in vivo when the cell has to cope with specific DNA lesions, such as N 2 -dG.

show abstract

Section: Discussionsupporting

confidence: 53%

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

Wiltrout¹,

Walker

2011

Genetics

View full text Add to dashboard Cite

show abstract

“…As expected for mutations that originate during G0, however, the frequency of two-strand mutations was unchanged in an mms2D or ubc13D strain. The major role of Rad5 is as the E3 ubiquitin ligase for the Ubc13-Mms2 E2 ubiquitin conjugase, but Rad5 also has been implicated in the Pol z bypass of some lesions (Gangavarapu et al 2006;Pagès et al 2008). In the ade2 adeX system used here, two-strand mutations were dependent on Rad5 as well as on Rad18.…”

Section: Discussionmentioning

confidence: 91%

“…As noted above, Rad5 is required for polyubiquitination of PCNA and hence is involved primarily in the error-free, template-switch pathway. Although loss of the error-free bypass pathway in dividing cells typically is associated with a mutator phenotype, Rad5 has been implicated in Pol z-dependent bypass of spontaneous lesions (Liefshitz et al 1998;Cejka et al 2001;Minesinger and Jinks-Robertson 2005) and of UV-lesion bypass in a gapped-plasmid transformation assay (Gangavarapu et al 2006;Pagès et al 2008). We observed no UV-induced mutagenesis in a rad5D mutant (irradiated to unirradiated sample comparison, P % 0.50), indicating that Rad5, like Rad18, is required for UVinduced mutagenesis in nondividing cells.…”

mentioning

confidence: 99%

The Mechanism of Nucleotide Excision Repair-Mediated UV-Induced Mutagenesis in Nonproliferating Cells

Kozmin

Jinks-Robertson

2013

Genetics

View full text Add to dashboard Cite

Following the irradiation of nondividing yeast cells with ultraviolet (UV) light, most induced mutations are inherited by both daughter cells, indicating that complementary changes are introduced into both strands of duplex DNA prior to replication. Early analyses demonstrated that such two-strand mutations depend on functional nucleotide excision repair (NER), but the molecular mechanism of this unique type of mutagenesis has not been further explored. In the experiments reported here, an ade2 adeX colonycolor system was used to examine the genetic control of UV-induced mutagenesis in nondividing cultures of Saccharomyces cerevisiae. We confirmed a strong suppression of two-strand mutagenesis in NER-deficient backgrounds and demonstrated that neither mismatch repair nor interstrand crosslink repair affects the production of these mutations. By contrast, proteins involved in the error-prone bypass of DNA damage (Rev3, Rev1, PCNA, Rad18, Pol32, and Rad5) and in the early steps of the DNA-damage checkpoint response (Rad17, Mec3, Ddc1, Mec1, and Rad9) were required for the production of two-strand mutations. There was no involvement, however, for the Pol h translesion synthesis DNA polymerase, the Mms2-Ubc13 postreplication repair complex, downstream DNA-damage checkpoint factors (Rad53, Chk1, and Dun1), or the Exo1 exonuclease. Our data support models in which UV-induced mutagenesis in nondividing cells occurs during the Pol z-dependent filling of lesion-containing, NER-generated gaps. The requirement for specific DNA-damage checkpoint proteins suggests roles in recruiting and/or activating factors required to fill such gaps. MUTAGENESIS associated with induced DNA damage is generally considered to occur during S phase, when polymerase-blocking lesions are bypassed in an error-prone manner by a translesion synthesis (TLS) DNA polymerase. In the yeast Saccharomyces cerevisiae, ultraviolet (UV)-induced mutagenesis is dependent on Pol z (Rev3-Rev7) and Rev1, with rev1, rev3, or rev7 mutants exhibiting a "reversionless" phenotype in response to UV irradiation (Lawrence 2002). Just as induced mutagenesis is considered to be a consequence of error-prone TLS, nucleotide excision repair (NER) is thought to counteract such mutagenesis by removing UV-induced lesions before they can be encountered during DNA replication.Although NER is generally considered to be an error-free, mutation-avoidance mechanism, data obtained .30 years ago demonstrated a pro-mutation role of NER in the UVinduced mutagenesis that occurs in nondividing yeast cells (Eckardt and Haynes 1977;James and Kilbey 1977;James et al. 1978;Eckardt et al. 1980). Either the induction of recessive lethal mutations in diploid strains (James and Kilbey 1977;James et al. 1978) or the induction of forward mutations in the de novo adenine biosynthetic pathway in haploid strains was monitored (Eckardt and Haynes 1977;Eckardt et al. 1980). Despite the differences in assay systems used, the authors reached the same conclusions. First, UVinduced mutations in nondi...

show abstract

“…However, subsequent results from analyses of the role of Rad5 in mutagenesis appeared to depend on the reporter allele used (Lawrence and Christensen, 1978). Recently, the ability to analyse the bypass of specific lesions at a known site in plasmids has allowed an unambiguous demonstration of the role of Rad5 in TLS in both S. cerevisiae (Pages et al, 2008;Zhang and Lawrence, 2005) and S. pombe (Coulon et al, 2010). Intriguingly, this role appears to be independent of Mms2 and Ubc13 in S. cerevisiae and, furthermore, does not require either the helicase or ubiquitin ligase activity of Rad5 (Pages et al, 2008), suggesting that Rad5 has a structural role.…”

Section: Crosstalk Between Template Switch and Tlsmentioning

confidence: 99%

Competition, collaboration and coordination – determining how cells bypass DNA damage

Sale

2012

Journal of Cell Science

View full text Add to dashboard Cite

SummaryCells must overcome replication blocks that might otherwise lead to genomic instability or cell death. Classical genetic experiments have identified a series of mechanisms that cells use to replicate damaged DNA: translesion synthesis, template switching and homologous recombination. In translesion synthesis, DNA lesions are replicated directly by specialised DNA polymerases, a potentially error-prone approach. Template switching and homologous recombination use an alternative undamaged template to allow the replicative polymerases to bypass DNA lesions and, hence, are generally error free. Classically, these pathways have been viewed as alternatives, competing to ensure replication of damaged DNA templates is completed. However, this view of a series of static pathways has been blurred by recent work using a combination of genetic approaches and methodology for examining the physical intermediates of bypass reactions. These studies have revealed a much more dynamic interaction between the pathways than was initially appreciated. In this Commentary, I argue that it might be more helpful to start thinking of lesion-bypass mechanisms in terms of a series of dynamically assembled 'modules', often comprising factors from different classical pathways, whose deployment is crucially dependent on the context in which the bypass event takes place.

show abstract

Requirement of Rad5 for DNA Polymerase ζ-Dependent Translesion Synthesis in Saccharomyces cerevisiae

Cited by 72 publications

References 50 publications

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

The DNA Polymerase Activity ofSaccharomyces cerevisiaeRev1 is Biologically Significant

The Mechanism of Nucleotide Excision Repair-Mediated UV-Induced Mutagenesis in Nonproliferating Cells

Competition, collaboration and coordination – determining how cells bypass DNA damage

Contact Info

Product

Resources

About