An essential role for dNTP homeostasis following CDK-induced replication stress

Pai, Chen Chun; Hsu, Kuo–Feng; Durley, Samuel C.; Keszthelyi, Andrea; Kearsey, Stephen; Rallis, Charalampos; Folkes, Lisa K.; Deegan, Rachel S.; Wilkins, Sarah E.; Pfister, Sophia X.; León, Nagore de; Schofield, Christopher J.; Bähler, Jürg; Carr, Antony; Humphrey, Timothy C.

doi:10.1242/jcs.226969

Cited by 15 publications

(10 citation statements)

References 67 publications

(90 reference statements)

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“…Aberrant Cdc2 activity is known to cause DNA damage in S. pombe cells (Caspari et al, 2002;Hills and Diffley, 2014;Deng et al, 2019) and this DNA damage may be the reason for the observed changes in Spc1 activity in mutants with aberrantly high Cdc2 activity. It has been reported that the presence of a hyperactive Cdc2 in Δwee1 cells causes dNTP depletion and replication stress (Pai et al, 2019). In combination with our results, this pointed to the possibility that the observed Spc1 activation in our experiments might be a response to replication stress.…”

Section: Replication Stress Caused By Cdc2 Hyperactivation Might Be Asupporting

confidence: 88%

Communication between Cyclin dependent kinase Cdc2 and the Wis1-Spc1 MAPK pathway determines mitotic timing in Schizosaccharomyces pombe

2020

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Checkpoint activation and gene expression modulation represent key determinants of cellular survival in adverse conditions. The former is regulated by cyclin-dependent kinases (CDKs) while the latter can be controlled by mitogen-activated protein kinases (MAPKs). Association between cell-cycle progression and MAPK-dependent gene expression exists in cells growing in optimal environments. While MAPK-mediated regulation of the cell cycle is well characterised, the reciprocal influence of mitotic CDK on stress response is not well studied. We present evidence that CDK activity can regulate the extent of MAPK activation in Schizosaccharomyces pombe cells. We show that increasing or decreasing mitotic CDK (Cdc2) activity in S. pombe cells can affect the activation of stress responsive MAPK (Spc1) even in the absence of stress stimuli. Our results indicate that the strong correlation between Cdc2 activity and Spc1 MAPK-activity in S. pombe is important in regulating mitotic timing. This article has an associated First Person interview with the first author of the paper.

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Section: Replication Stress Caused By Cdc2 Hyperactivation Might Be Asupporting

confidence: 88%

Communication between Cyclin dependent kinase Cdc2 and the Wis1-Spc1 MAPK pathway determines mitotic timing in Schizosaccharomyces pombe

2020

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“…dNTP levels were determined as previously described [76]. S. pombe pol2 variant strains were constructed as described previously [77].…”

Section: S Pombe Methodsmentioning

confidence: 99%

Expression of the cancer-associated DNA polymerase ε P286R in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

et al. 2021

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Somatic and germline mutations in the proofreading domain of the replicative DNA polymerase ε (POLE-exonuclease domain mutations, POLE-EDMs) are frequently found in colorectal and endometrial cancers and, occasionally, in other tumours. POLE-associated cancers typically display hypermutation, and a unique mutational signature, with a predominance of C > A transversions in the context TCT and C > T transitions in the context TCG. To understand better the contribution of hypermutagenesis to tumour development, we have modelled the most recurrent POLE-EDM (POLE-P286R) in Schizosaccharomyces pombe. Whole-genome sequencing analysis revealed that the corresponding pol2-P287R allele also has a strong mutator effect in vivo, with a high frequency of base substitutions and relatively few indel mutations. The mutations are equally distributed across different genomic regions, but in the immediate vicinity there is an asymmetry in AT frequency. The most abundant base-pair changes are TCT > TAT transversions and, in contrast to human mutations, TCG > TTG transitions are not elevated, likely due to the absence of cytosine methylation in fission yeast. The pol2-P287R variant has an increased sensitivity to elevated dNTP levels and DNA damaging agents, and shows reduced viability on depletion of the Pfh1 helicase. In addition, S phase is aberrant and RPA foci are elevated, suggestive of ssDNA or DNA damage, and the pol2-P287R mutation is synthetically lethal with rad3 inactivation, indicative of checkpoint activation. Significantly, deletion of genes encoding some translesion synthesis polymerases, most notably Pol κ, partially suppresses pol2-P287R hypermutation, indicating that polymerase switching contributes to this phenotype.

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“…Fluorescence microscopy was carried out as previously described [66], except cells were not fixed. dNTP levels were determined as previously described [67].…”

Section: S Pombe Methodsmentioning

confidence: 99%

Expression of the cancer-associated DNA polymerase ε P286R in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

Soriano

Vázquez

León

et al. 2021

Preprint

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Somatic mutations in the proofreading domain of the replicative DNA polymerase ϵ ( POLE- exonuclease domain mutations, POLE -EDMs) are frequently found in colorectal and endometrial cancers and, occasionally, in other tumours. POLE-associated cancers typically display hypermutation, microsatellite stability and a unique mutational signature, with a predominance of C > A transversions in the context TCT. To understand better the contribution of hypermutagenesis to tumour development, we have modelled the most recurrent POLE -EDM ( POLE-P286R ) in Schizosaccharomyces pombe . Whole-genome sequencing analysis revealed that the corresponding pol2-P287R allele also has a strong mutator effect in vivo, with a high frequency of base substitutions and relatively few frameshift mutations. The mutations are equally distributed across different genomic regions, but they occur within an AT-rich context. The most abundant base-pair changes are T C T > T A T transversions and, in contrast to human mutations, T C G > T T G transitions are not elevated, likely due to the absence of cytosine methylation in fission yeast. The pol2-P287R variant has an increased sensitivity to elevated dNTP levels and DNA damaging agents, and shows reduced viability on depletion of the Pfh1 helicase. In addition, S phase is aberrant and RPA foci are elevated, suggestive of persistent ssDNA or DNA damage, and the pol2-P287R mutation is synthetically lethal with rad3 inactivation , indicative of checkpoint activation. Significantly, deletion of genes encoding some translesion synthesis polymerases, most notably Pol κ, partially suppresses pol2-P287R hypermutation, indicating that polymerase switching contributes to this phenotype.

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An essential role for dNTP homeostasis following CDK-induced replication stress

Cited by 15 publications

References 67 publications

Communication between Cyclin dependent kinase Cdc2 and the Wis1-Spc1 MAPK pathway determines mitotic timing in Schizosaccharomyces pombe

Communication between Cyclin dependent kinase Cdc2 and the Wis1-Spc1 MAPK pathway determines mitotic timing in Schizosaccharomyces pombe

Expression of the cancer-associated DNA polymerase ε P286R in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

Expression of the cancer-associated DNA polymerase ε P286R in fission yeast leads to translesion synthesis polymerase dependent hypermutation and defective DNA replication

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