The Fission Yeast UVDR DNA Repair Pathway Is Inducible

Davey, Scott; Nass, Monica; Ferrer, Jasmine V.; Sidik, Khalifah; Eisenberger, Andrew; Mitchell, David L.; Freyer, Greg A.

doi:10.1093/nar/25.5.1002

Cited by 25 publications

(26 citation statements)

References 34 publications

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“…Also, the extents of induction were comparable; when normalized to control leu1 ϩ mRNA, and compared to uninduced mRNA levels, the maximal observed uve1 ϩ mRNA inductions were 3.4-fold in wild type and 3.7-fold in rad9 mutant cells. This indicates that the elevation of UVDE activity in rad9 mutants is the result of a posttranscriptional regulation which functions in addition to the previously reported transcriptional regulation of uve1 ϩ following UV treatment (7).…”

Section: Rad12supporting

confidence: 59%

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Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Davey

Han

Ramer³

et al. 1998

Molecular and Cellular Biology

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The human BLM gene is a member of the Escherichia coli recQ helicase family, which includes the Saccharomyces cerevisiae SGS1 and human WRN genes. Defects in BLM are responsible for the human disease Bloom's syndrome, which is characterized in part by genomic instability and a high incidence of cancer. Here we describe the cloning of rad12 ؉ , which is the fission yeast homolog of BLM and is identical to the recently reported rhq1؉ gene. We showed that rad12 null cells are sensitive to DNA damage induced by UV light and ␥ radiation, as well as to the DNA synthesis inhibitor hydroxyurea. Overexpression of the wild-type rad12 ؉ gene also leads to sensitivity to these agents and to defects associated with the loss of the S-phase and G 2 -phase checkpoint control. We showed genetically and biochemically that rad12 ؉ acts upstream from rad9 ؉ , one of the fission yeast G 2 checkpoint control genes, in regulating exit from the S-phase checkpoint. The physical chromosome segregation defects seen in rad12 null cells combined with the checkpoint regulation defect seen in the rad12 ؉ overproducer implicate rad12 ؉ as a key coupler of chromosomal integrity with cell cycle progression.The fission yeast Schizosaccharomyces pombe undergoes a dose-dependent G 2 delay in response to DNA damage caused by radiation (1, 27). Cells remain arrested at this G 2 checkpoint while DNA damage is repaired, then enter mitosis and resume progression through the cell cycle. Six checkpoint rad genes have been identified in S. pombe: rad1 ϩ , rad3ϩ , rad26 ϩ , and hus1 ϩ (1, 2, 9, 27). Mutations in any of these genes result in almost identical phenotypes. The mutant strains are all sensitive to radiation and to agents which transiently inhibit DNA replication, and all lack the G 2 checkpoint, preventing mitotic entry in the presence of such damage (1, 2, 9, 27). While little is known about G 2 checkpoint genes in humans, homologs of S. pombe rad3 ϩ (4, 5) and rad9 ϩ (19) have recently been isolated. Recent studies have implicated cell cycle gene mutations in causing cancer. It is likely that mutations in genes regulating the G 1 and G 2 checkpoints also have the potential to increase cancer susceptibility.The phenomenon of genomic instability has been firmly established as a crucial step in the genesis of cancer (14,31,33). While genomic instability is seen as a step in the progression of cancer cells, there are also several genetic diseases that lead to genomic instability and, ultimately, to cancer. Among these diseases is the rare autosomal recessive disorder Bloom's syndrome (12). The defective gene in Bloom's syndrome, BLM, encodes a putative DNA helicase that is homologous to the Escherichia coli RecQ and Saccharomyces cerevisiae Sgs1 helicases (11,34). While the BLM and SGS1 proteins contain centrally located helicase domains that are homologous to RecQ, they are much larger than RecQ and show homology, both 5Ј and 3Ј, to the helicase core (26, 35, 37).We describe here the cloning of the fission yeast rad12 ϩ gene, which encodes a helic...

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Section: Rad12supporting

confidence: 59%

“…Whole-cell extracts were prepared from 10 9 S. pombe cells as described previously (7). The protein concentrations of extracts prepared this way were approximately 20 to 30 mg/ml.…”

Section: S Pombe Genetic Manipulations S Pombe Was Cultured By Stamentioning

confidence: 99%

Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Davey

Han

Ramer³

et al. 1998

Molecular and Cellular Biology

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“…studies provide interesting implications for the role of the polyalanine tract in the control of rhp51 ϩ transcription. Several S. pombe genes, including uvi15 ϩ , uvi31 ϩ , UVDE, and rhp16 ϩ , are DNA damage-inducible genes (5,14). Of these, uvi31 ϩ , UVDE, and rhp16 ϩ contain a putative sequence homologous to DRE rhp51 ϩ (5, 14, 37), suggesting the presence of a DDR controlled by DRE rhp51 ϩ.…”

Section: Discussionmentioning

confidence: 99%

Rdp1, a Novel Zinc Finger Protein, Regulates the DNA Damage Response of rhp51⁺ from Schizosaccharomyces pombe

Shim

Jang

Lim

et al. 2000

Molecular and Cellular Biology

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The Schizosaccharomyces pombe DNA repair gene rhp51؉ encodes a RecA-like protein with the DNAdependent ATPase activity required for homologous recombination. The level of the rhp51 ؉ transcript is increased by a variety of DNA-damaging agents. Its promoter has two cis-acting DNA damage-responsive elements (DREs) responsible for DNA damage inducibility. Here we report identification of Rdp1, which regulates rhp51؉ expression through the DRE of rhp51 ؉ . The protein contains a zinc finger and a polyalanine tract similar to ones previously implicated in DNA binding and transactivation or repression, respectively. In vitro footprinting and competitive binding assays indicate that the core consensus sequences (NGG/TTG/A) of DRE are crucial for the binding of Rdp1. Mutations of both DRE1 and DRE2 affected the damage-induced expression of rhp51 ؉ , indicating that both DREs are required for transcriptional activation. In addition, mutations in the DREs significantly reduced survival rates after exposure to DNA-damaging agents, demonstrating that the damage response of rhp51 ؉ enhances the cellular repair capacity. Surprisingly, haploid cells containing a complete rdp1 deletion could not be recovered, indicating that rdp1؉ is essential for cell viability and implying the existence of other target genes. Furthermore, the DNA damage-dependent expression of rhp51 ؉ was significantly reduced in checkpoint mutants, raising the possibility that Rdp1 may mediate damage checkpoint-dependent transcription of rhp51All organisms have developed defense mechanisms to respond to genotoxic materials causing genetic injury. One response to DNA damage or DNA synthesis inhibition is to delay the cell cycle by blocking DNA replication and/or mitotic division. Another is the transcriptional induction of several genes whose products may contribute to DNA repair capacity (22).During past decades, such damage-inducible genes have been identified and partially characterized for bacteria, yeasts, and higher eukaryotes (22). In particular, a large number of genes are induced in response to DNA damage and/or inhibition of DNA replication in Saccharomyces cerevisiae (22,49). These include RNR2 (which encodes a small subunit of ribonucleotide reductase [19,28] [12]), which are involved in DNA repair. However, the biological significance of the transcriptional induction of these genes has been uncovered only recently. A study has revealed that Dun1p serine/threonine protein kinase is involved in the transcriptional activation of RNR2 and has delineated a pathway by which the damage signal is transduced to a checkpoint and transcription response apparatus (65). The repressor protein Crt1p was found to bind to X boxes on the RNR2 and RNR3 promoters and to mediate repression of the genes by cooperating with the Tup1p-Ssn6p corepressor. DNA damage-induced hyperphosphorylation of Crt1p enables the protein to dissociate from X boxes, which leads to derepression of RNR2 transcription. This dissociation is also dependent on the MEC1-RAD53-DUN1 damage-signal...

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Section: Introductionmentioning

confidence: 99%

“…In lower eukaryotes, studies with the budding yeast Saccharomyces cerevisiae have identified a large number of damageinduced genes, including many involved in DNA metabolism and DNA repair (Aboussekhra et al, 1996;Gasch et al, 2001). In the distantly related fission yeast Schizosaccharomyces pombe, rhp51 ϩ , uvi15 ϩ , uvi31 ϩ , UVDE ϩ , and rhp16 ϩ have been shown to be DNA damage inducible (Bang et al, 1996;Davey et al, 1997;Shim et al, 2000).DNA damage checkpoint pathways function to delay the eukaryotic cell cycle in response to DNA damage, thus providing an opportunity for DNA repair. Central to the DNA damage checkpoint pathway are two highly conserved phosphoinositol-related kinases, Ataxia telangiectasia mutated (ATM) and ATM and Rad3-related (ATR) and their yeast homologs SpTel1 and SpRad3 in S. pombe, and ScTel1 and ScMec1 in S. cerevisiae.…”

mentioning

confidence: 99%

Global Gene Expression Responses of Fission Yeast to Ionizing Radiation

Watson

Mata

Bähler

et al. 2004

MBoC

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A coordinated transcriptional response to DNA-damaging agents is required to maintain genome stability. We have examined the global gene expression responses of the fission yeast Schizosaccharomyces pombe to ionizing radiation (IR) by using DNA microarrays. We identified ϳ200 genes whose transcript levels were significantly altered at least twofold in response to 500 Gy of gamma IR in a temporally defined manner. The majority of induced genes were core environmental stress response genes, whereas the remaining genes define a transcriptional response to DNA damage in fission yeast. Surprisingly, few DNA repair and checkpoint genes were transcriptionally modulated in response to IR. We define a role for the stress-activated mitogen-activated protein kinase Sty1/Spc1 and the DNA damage checkpoint kinase Rad3 in regulating core environmental stress response genes and IR-specific response genes, both independently and in concert. These findings suggest a complex network of regulatory pathways coordinate gene expression responses to IR in eukaryotes. INTRODUCTIONExposure of cells to DNA-damaging agents such as ionizing radiation (IR) poses a significant threat to genome stability. Cells have therefore evolved complex response mechanisms to maintain genetic integrity after DNA damage. These include cell cycle delay, repair of DNA damage, transcriptional responses, and programmed cell death. Because disruption of any one of these DNA damage responses can lead to genomic instability and cancer, a comprehensive understanding of the individual components and their regulation is crucial.It is well established that many physiological responses to DNA damage are regulated at the level of gene expression. In Escherichia coli, the SOS response induces the expression of a network of genes after DNA damage through the regulatory LexA and RecA proteins (Friedberg et al., 1995). In lower eukaryotes, studies with the budding yeast Saccharomyces cerevisiae have identified a large number of damageinduced genes, including many involved in DNA metabolism and DNA repair (Aboussekhra et al., 1996;Gasch et al., 2001). In the distantly related fission yeast Schizosaccharomyces pombe, rhp51 ϩ , uvi15 ϩ , uvi31 ϩ , UVDE ϩ , and rhp16 ϩ have been shown to be DNA damage inducible (Bang et al., 1996;Davey et al., 1997;Shim et al., 2000).DNA damage checkpoint pathways function to delay the eukaryotic cell cycle in response to DNA damage, thus providing an opportunity for DNA repair. Central to the DNA damage checkpoint pathway are two highly conserved phosphoinositol-related kinases, Ataxia telangiectasia mutated (ATM) and ATM and Rad3-related (ATR) and their yeast homologs SpTel1 and SpRad3 in S. pombe, and ScTel1 and ScMec1 in S. cerevisiae. Activation of ATM and ATR kinases by DNA damage leads to cell cycle arrest through a number of downstream effector molecules including the effector kinases CHK1/SpChk1/ScChk1 and CHK2/SpCds1/ ScRad53 (for review, see Nyberg et al., 2002). In addition to regulating the cell cycle, DNA damage checkpoints also...

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The Fission Yeast UVDR DNA Repair Pathway Is Inducible

Cited by 25 publications

References 34 publications

Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Rdp1, a Novel Zinc Finger Protein, Regulates the DNA Damage Response of rhp51⁺ from Schizosaccharomyces pombe

Global Gene Expression Responses of Fission Yeast to Ionizing Radiation

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The Fission Yeast UVDR DNA Repair Pathway Is Inducible

Cited by 25 publications

References 34 publications

Fission Yeastrad12+Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Fission Yeastrad12+Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Rdp1, a Novel Zinc Finger Protein, Regulates the DNA Damage Response of rhp51+ from Schizosaccharomyces pombe

Global Gene Expression Responses of Fission Yeast to Ionizing Radiation

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Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Fission Yeastrad12⁺Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

Rdp1, a Novel Zinc Finger Protein, Regulates the DNA Damage Response of rhp51⁺ from Schizosaccharomyces pombe