Activation of Checkpoint Kinase Chk1 by Reactive Oxygen Species Resulting from Disruption of wat1/pop3 in Schizosaccharomyces pombe.
DNA double-strand breaks are critical lesions that can lead to chromosomal aberrations and genomic instability. In response to DNA damage, Chk1, a serine/threonine kinase, is responsible for cell cycle arrest to prevent damaged cells from progressing through the cell cycle. Here, we report that the disruption of wat1, a WD repeat-containing protein, leads to the phosphorylation of Chk1. The double-deletion of chk1 and wat1 had a grave effect on the survival of fission yeast cells, and the spontaneous recombination rate was also high upon double-deletion of wat1 and chk1, as compared to the single-mutant. In the absence of wat1, the cells exhibited a high level of nuclear fragmentation that resulted in the accumulation of Rad22 yellow fluorescent protein foci. Furthermore, we show that wat1 is required for the regulation of the oxidative stress response. We observed elevated levels of reactive oxygen species (ROS) generation in wat1-null mutant that led to a high degree of propidium iodide staining at nonpermissive temperature. Based on the results presented here, we hypothesize that ROS production in wat1-null mutant cells generates DNA fragmentation that could trigger a checkpoint response and that, in the absence of checkpoint kinase Chk1, the cells exhibit severe growth defects leading to a synthetic lethal phenotype.
Molecular and Structural Biology Division, Council of Scientific & Industrial Research-Central Drug Research Institute, Lucknow 226031, Uttar Pradesh, India.
Nafees
http://orcid.org/0000-0002-8944-1819
N
Ahamad
Molecular and Structural Biology Division, Council of Scientific & Industrial Research-Central Drug Research Institute, Lucknow 226031, Uttar Pradesh, India.
Sumit Kumar
http://orcid.org/0000-0003-3507-7780
SK
Verma
Molecular and Structural Biology Division, Council of Scientific & Industrial Research-Central Drug Research Institute, Lucknow 226031, Uttar Pradesh, India shakil_ahmed@cdri.res.in.
Shakil
S
Ahmed
D017209:Apoptosis; D000071877:Checkpoint Kinase 1 / Q000235:genetics / Q000378:metabolism*; D053938:DNA Fragmentation; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism*
C105525:Pop3 protein, S pombe; D017382:Reactive Oxygen Species; D029702:Schizosaccharomyces pombe Proteins; D000071877:Checkpoint Kinase 1; C000605873:Chk1 protein, S pombe
Chk1; Wat1/Pop3; apoptosis; cell death; reactive oxygen species
Environ Health Perspect. 1994 Dec;102 Suppl 10:5-12
7705305
PLoS One. 2014 Feb 21;9(2):e89587
24586893
J Cell Sci. 2006 Jan 1;119(Pt 1):124-31
16371652
Yeast. 1999 Jul;15(10A):821-8
10407262
Free Radic Biol Med. 2001 Dec 15;31(12):1624-32
11744337
Cytometry. 1994 Aug 1;16(4):305-12
7527314
Biochimie. 2002 Feb-Mar;84(2-3):131-41
12022944
Yeast. 1998 Jul;14 (10 ):943-51
9717240
J Cell Biol. 1999 May 17;145(4):757-67
10330404
Proc Natl Acad Sci U S A. 2012 Jan 31;109 (5):1536-41
22307609
Curr Opin Cell Biol. 2004 Dec;16(6):629-33
15530773
Nature. 1993 May 27;363(6427):368-71
8497322
Genes Cells. 2007 Dec;12(12):1357-70
18076573
DNA Repair (Amst). 2002 Dec 5;1(12 ):983-94
12531008
Genetics. 1997 Dec;147(4):1569-84
9409822
Proc Natl Acad Sci U S A. 1986 Apr;83(7):2162-6
3083416
Mol Cell. 2013 Sep 26;51(6):829-39
24035500
PLoS One. 2015 Mar 30;10 (3):e0124063
25822347
Curr Genet. 2000 Oct;38(3):113-25
11057444
Redox Biol. 2014 Jan 09;2:457-65
24624335
J Biol Chem. 2006 Nov 3;281(44):33000-7
16959779
Mol Biol Cell. 2002 May;13(5):1536-49
12006651
Nat Rev Cancer. 2003 Mar;3(3):155-68
12612651
Genes Dev. 1997 Feb 15;11(4):504-11
9042863
J Cell Sci. 2001 Aug;114(Pt 16):2911-20
11686295
J Bioenerg Biomembr. 2004 Aug;36(4):341-6
15377869
Free Radic Biol Med. 2003 Feb 15;34(4):478-88
12566073
Mol Biol Cell. 1994 Feb;5(2):147-60
8019001
Mutat Res. 2002 Jan 15;513(1-2):69-74
11719091
J Biol Chem. 2003 Apr 25;278(17):15136-41
12569108
Science. 1996 Jan 19;271(5247):353-6
8553071
J Biol Chem. 2007 Mar 9;282(10 ):7125-36
17215245
Annu Rev Genet. 2002;36:617-56
12429704
Methods Enzymol. 1991;194:795-823
2005825
Curr Biol. 2003 Dec 2;13(23):R904-5
14654013
J Cell Sci. 2005 Aug 1;118(Pt 15):3543-53
16079294
Mol Microbiol. 2012 Dec;86(5):1246-61
23062268
Nature. 2013 May 9;497(7448):217-23
23636326
Front Oncol. 2012 Jun 20;2:64
22737670
Proc Natl Acad Sci U S A. 2013 Jun 25;110(26):10592-7
23754435
Annu Rev Genet. 2007;41:237-80
17630848
Nature. 1994 Sep 22;371(6495):297-300
8090199
Mol Gen Genet. 1997 Mar 26;254(2):127-38
9108274