Mammalian Fbh1 is important to restore normal mitotic progression following decatenation stress.
We have addressed the role of the F-box helicase 1 (Fbh1) protein during genome maintenance in mammalian cells. For this, we generated two mouse embryonic stem cell lines deficient for Fbh1: one with a homozygous deletion of the N-terminal F-box domain (Fbh1(f/f)), and the other with a homozygous disruption (Fbh1(-/-)). Consistent with previous reports of Fbh1-deficiency in vertebrate cells, we found that Fbh1(-/-) cells show a moderate increase in Rad51 localization to DNA damage, but no clear defect in chromosome break repair. In contrast, we found that Fbh1(f/f) cells show a decrease in Rad51 localization to DNA damage and increased cytoplasmic localization of Rad51. However, these Fbh1(f/f) cells show no clear defects in chromosome break repair. Since some Rad51 partners and F-box-associated proteins (Skp1-Cul1) have been implicated in progression through mitosis, we considered whether Fbh1 might play a role in this process. To test this hypothesis, we disrupted mitosis using catalytic topoisomerase II inhibitors (bisdioxopiperazines), which inhibit chromosome decatenation. We found that both Fbh1(f/f) and Fbh1(-/-) cells show hypersensitivity to topoisomerase II catalytic inhibitors, even though the degree of decatenation stress was not affected. Furthermore, following topoisomerase II catalytic inhibition, both Fbh1-deficient cell lines show substantial defects in anaphase separation of chromosomes. These results indicate that Fbh1 is important for restoration of normal mitotic progression following decatenation stress.
Laulier
Corentin
C
Department of Cancer Biology, Division of Radiation Biology, Beckman Research Institute of the City of Hope, 1500 E Duarte Rd., Duarte, CA 91010, USA.
Cheng
Anita
A
Huang
Nick
N
Stark
Jeremy M
JM
D000705:Anaphase / Q000187:drug effects / Q000235:genetics; D000818:Animals; D055162:Biocatalysis; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D019457:Chromosome Breakage / Q000187:drug effects; D004265:DNA Helicases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D043864:DNA, Catenated / Q000378:metabolism; D054659:Diketopiperazines; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D044783:F-Box Proteins; D017353:Gene Deletion; D006720:Homozygote; D006801:Humans; D051379:Mice; D008938:Mitosis / Q000502:physiology*; D010879:Piperazines / Q000494:pharmacology; D021381:Protein Transport; D051135:Rad51 Recombinase / Q000378:metabolism; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics*; D059005:Topoisomerase II Inhibitors
D043864:DNA, Catenated; D054659:Diketopiperazines; D044783:F-Box Proteins; D010879:Piperazines; D059005:Topoisomerase II Inhibitors; C070899:4,4'-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione; D051135:Rad51 Recombinase; D004265:DNA Helicases; C553250:Fbh1 protein, mouse; D004250:DNA Topoisomerases, Type II
J Cell Biol. 1994 Sep;126(6):1341-51
8089169
Cancer Res. 1991 Sep 15;51(18):4903-8
1654204
Nat Rev Mol Cell Biol. 2004 Sep;5(9):739-51
15340381
Mol Cell. 2004 Sep 24;15(6):977-90
15383286
Cancer Res. 1991 Sep 15;51(18):4909-16
1654205
J Cell Biol. 2009 Sep 7;186(5):655-63
19736316
Nature. 2001 Nov 1;414(6859):105-11
11689955
Proc Natl Acad Sci U S A. 2001 Oct 9;98(21):12044-9
11593014
Mol Cell Biol. 2003 Jan;23(2):733-43
12509470
Curr Biol. 2009 Jul 14;19(13):1075-85
19540122
PLoS Genet. 2008 Jun 27;4(6):e1000110
18584027
Mol Cell Biol. 2007 Nov;27(21):7439-50
17724085
Cell. 2007 Jan 12;128(1):101-14
17218258
J Biol Chem. 2002 Jul 5;277(27):24530-7
11956208
EMBO J. 2007 Jul 25;26(14):3397-409
17599064
Nat Rev Cancer. 2005 Oct;5(10):773-85
16195750
J Clin Oncol. 2009 Jan 1;27(1):127-45
19018081
Cancer Res. 2009 Aug 15;69(16):6668-75
19679553
Mol Cell. 2001 Feb;7(2):263-72
11239455
Nature. 1994 Dec 1;372(6505):467-70
7984241
Genes Dev. 1999 Sep 1;13(17):2242-57
10485847
Mol Cell Biol. 2005 Sep;25(18):8074-83
16135799
Biochem Pharmacol. 1995 Sep 28;50(7):953-8
7575679
Mutat Res. 2006 Aug 30;600(1-2):58-66
16822529
Nat Rev Cancer. 2009 May;9(5):327-37
19377505
Development. 2006 Feb;133(4):663-73
16421190
Cancer Res. 1994 Feb 15;54(4):1042-8
8313360
Curr Biol. 1998 Mar 26;8(7):377-85
9545195
Nucleic Acids Res. 2004 Apr 26;32(8):2287-97
15118074
Nat Rev Cancer. 2009 May;9(5):338-50
19377506
Mol Cell. 2008 Jun 20;30(6):790-802
18570880
J Biol Chem. 2003 Sep 19;278(38):35897-902
12842886
Curr Biol. 2009 Dec 1;19(22):1907-11
19836238
Mol Cell Biol. 2009 Sep;29(17):4742-56
19546232
Cell. 1997 Oct 17;91(2):209-19
9346238
Mol Cell Biol. 2005 Sep;25(18):8084-96
16135800
J Cell Sci. 2008 Jan;121 Suppl 1:1-84
18089652
Br J Cancer. 2007 Jan 29;96(2):201-5
17211475
J Biol Chem. 2003 Oct 31;278(44):42729-32
12912992
Cancer Res. 2009 Feb 1;69(3):1125-34
19176389
Nat Struct Mol Biol. 2005 Jul;12(7):589-93
15965487
J Cell Sci. 1993 Jun;105 ( Pt 2):563-9
8408285
Mol Cell Biol. 2007 Apr;27(8):2812-20
17283053
Mutagenesis. 2003 May;18(3):221-33
12714687
J Cell Sci. 2007 Nov 15;120(Pt 22):3952-64
17956945
PLoS One. 2009;4(4):e5323
19390626
Science. 2004 Oct 29;306(5697):876-9
15375219
Cell. 2002 May 17;109(4):459-72
12086603
Cancer Cell. 2005 Dec;8(6):479-84
16338661
Genes Dev. 2004 Nov 1;18(21):2573-80
15520277