scholarly journals Monitoring Schizosaccharomyces pombe genome stress by visualizing end-binding protein Ku

Biology Open ◽  
2021 ◽  
Vol 10 (2) ◽  
Author(s):  
Chance E. Jones ◽  
Susan L. Forsburg
Keyword(s):  
Dna Damage ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Protein Complex ◽  
Genome Stability ◽  
Binding Protein ◽  
Dna Lesions ◽  
Live Cells ◽  
Dna Damage Checkpoint ◽  
Ku Protein

ABSTRACT Studies of genome stability have exploited visualization of fluorescently tagged proteins in live cells to characterize DNA damage, checkpoint, and repair responses. In this report, we describe a new tool for fission yeast, a tagged version of the end-binding protein Pku70 which is part of the KU protein complex. We compare Pku70 localization to other markers upon treatment to various genotoxins, and identify a unique pattern of distribution. Pku70 provides a new tool to define and characterize DNA lesions and the repair response.

scholarly journals Monitoring S. pombe genome stress by visualizing end-binding protein Ku

2020 ◽  
Author(s):  
Chance Jones ◽  
Susan L Forsburg
Keyword(s):  
Dna Damage ◽  
Fission Yeast ◽  
Protein Complex ◽  
Genome Stability ◽  
Binding Protein ◽  
Dna Lesions ◽  
Live Cells ◽  
Dna Damage Checkpoint ◽  
Unique Pattern ◽  
Ku Protein

AbstractStudies of genome stability have exploited visualization of fluorescently tagged proteins in live cells to characterize DNA damage, checkpoint, and repair responses. In this report, we describe a new tool for fission yeast, a tagged version of the end-binding protein Pku70 which is part of the KU protein complex. We compare Pku70 localization to other markers upon treatment to various genotoxins, and identify a unique pattern of distribution. Pku70 provides a new tool to define and characterize DNA lesions and the repair response.

scholarly journals The Fission Yeast Crb2/Chk1 Pathway Coordinates the DNA Damage and Spindle Checkpoint in Response to Replication Stress Induced by Topoisomerase I Inhibitor

2005 ◽  
Vol 25 (17) ◽  
pp. 7889-7899 ◽  
Author(s):  
Ada Collura ◽  
Joel Blaisonneau ◽  
Giuseppe Baldacci ◽  
Stefania Francesconi
Keyword(s):  
Dna Damage ◽  
Fission Yeast ◽  
Topoisomerase I ◽  
Genome Stability ◽  
Cell Cycle Progression ◽  
Spindle Assembly Checkpoint ◽  
Genetic Material ◽  
Spindle Checkpoint ◽  
Dna Damage Checkpoint ◽  
Replicative Stress

ABSTRACT Living organisms experience constant threats that challenge their genome stability. The DNA damage checkpoint pathway coordinates cell cycle progression with DNA repair when DNA is damaged, thus ensuring faithful transmission of the genome. The spindle assembly checkpoint inhibits chromosome segregation until all chromosomes are properly attached to the spindle, ensuring accurate partition of the genetic material. Both the DNA damage and spindle checkpoint pathways participate in genome integrity. However, no clear connection between these two pathways has been described. Here, we analyze mutants in the BRCT domains of fission yeast Crb2, which mediates Chk1 activation, and provide evidence for a novel function of the Chk1 pathway. When the Crb2 mutants experience damaged replication forks upon inhibition of the religation activity of topoisomerase I, the Chk1 DNA damage pathway induces sustained activation of the spindle checkpoint, which in turn delays metaphase-to-anaphase transition in a Mad2-dependent fashion. This new pathway enhances cell survival and genome stability when cells undergo replicative stress in the absence of a proficient G2/M DNA damage checkpoint.

scholarly journals Mechanism of Caffeine-Induced Checkpoint Override in Fission Yeast

2000 ◽  
Vol 20 (12) ◽  
pp. 4288-4294 ◽  
Author(s):  
Bettina A. Moser ◽  
Jean-Marc Brondello ◽  
Beth Baber-Furnari ◽  
Paul Russell
Keyword(s):  
Dna Damage ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Protein Kinases ◽  
Molecular Target ◽  
S Phase ◽  
Dna Damage Checkpoint ◽  
Mitotic Checkpoints ◽  
Direct Activation

ABSTRACT Mitotic checkpoints restrain the onset of mitosis (M) when DNA is incompletely replicated or damaged. These checkpoints are conserved between the fission yeast Schizosaccharomyces pombe and mammals. In both types of organisms, the methylxanthine caffeine overrides the synthesis (S)-M checkpoint that couples mitosis to completion of DNA S phase. The molecular target of caffeine was sought in fission yeast. Caffeine prevented activation of Cds1 and phosphorylation of Chk1, two protein kinases that enforce the S-M checkpoint triggered by hydroxyurea. Caffeine did not inhibit these kinases in vitro but did inhibit Rad3, a kinase that regulates Cds1 and Chk1. In accordance with this finding, caffeine also overrode the G2-M DNA damage checkpoint that requires Rad3 function. Rad3 coprecipitated with Cds1 expressed at endogenous amounts, a finding that supports the hypothesis that Rad3 is involved in direct activation of Cds1.

scholarly journals Roles of the Mitotic Inhibitors Wee1 and Mik1 in the G2 DNA Damage and Replication Checkpoints

2001 ◽  
Vol 21 (5) ◽  
pp. 1499-1508 ◽  
Author(s):  
Nicholas Rhind ◽  
Paul Russell
Keyword(s):  
Dna Damage ◽  
Dna Replication ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dna Damage Checkpoint ◽  
Mitotic Inhibitors

ABSTRACT The G2 DNA damage and DNA replication checkpoints in many organisms act through the inhibitory phosphorylation of Cdc2 on tyrosine-15. This phosphorylation is catalyzed by the Wee1/Mik1 family of kinases. However, the in vivo role of these kinases in checkpoint regulation has been unclear. We show that, in the fission yeastSchizosaccharomyces pombe, Mik1 is a target of both checkpoints and that the regulation of Mik1 is, on its own, sufficient to delay mitosis in response to the checkpoints. Mik1 appears to have two roles in the DNA damage checkpoint; one in the establishment of the checkpoint and another in its maintenance. In contrast, Wee1 does not appear to be involved in the establishment of either checkpoint.

scholarly journals PARP-1–dependent recruitment of cold-inducible RNA-binding protein promotes double-strand break repair and genome stability

2018 ◽  
Vol 115 (8) ◽  
pp. E1759-E1768 ◽  
Author(s):  
Jung-Kuei Chen ◽  
Wen-Ling Lin ◽  
Zhang Chen ◽  
Hung-wen Liu
Keyword(s):  
Dna Damage ◽  
Genome Stability ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Strand Break ◽  
Double Strand Break ◽  
Dsb Repair ◽  
Active Involvement ◽  
Parp 1

Maintenance of genome integrity is critical for both faithful propagation of genetic information and prevention of mutagenesis induced by various DNA damage events. Here we report cold-inducible RNA-binding protein (CIRBP) as a newly identified key regulator in DNA double-strand break (DSB) repair. On DNA damage, CIRBP temporarily accumulates at the damaged regions and is poly(ADP ribosyl)ated by poly(ADP ribose) polymerase-1 (PARP-1). Its dissociation from the sites of damage may depend on its phosphorylation status as mediated by phosphatidylinositol 3-kinase-related kinases. In the absence of CIRBP, cells showed reduced γH2AX, Rad51, and 53BP1 foci formation. Moreover, CIRBP-depleted cells exhibited impaired homologous recombination, impaired nonhomologous end-joining, increased micronuclei formation, and higher sensitivity to gamma irradiation, demonstrating the active involvement of CIRBP in DSB repair. Furthermore, CIRBP depleted cells exhibited defects in DNA damage-induced chromatin association of the MRN complex (Mre11, Rad50, and NBS1) and ATM kinase. CIRBP depletion also reduced phosphorylation of a variety of ATM substrate proteins and thus impaired the DNA damage response. Taken together, these results reveal a previously unrecognized role for CIRBP in DSB repair.

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