New Functions of Ctf18-RFC in Preserving Genome Stability outside Its Role in Sister Chromatid Cohesion
Expansion of DNA trinucleotide repeats causes at least 15 hereditary neurological diseases, and these repeats also undergo contraction and fragility. Current models to explain this genetic instability invoke erroneous DNA repair or aberrant replication. Here we show that CAG/CTG tracts are stabilized in Saccharomyces cerevisiae by the alternative clamp loader/unloader Ctf18-Dcc1-Ctf8-RFC complex (Ctf18-RFC). Mutants in Ctf18-RFC increased all three forms of triplet repeat instability—expansions, contractions, and fragility—with effect over a wide range of allele lengths from 20–155 repeats. Ctf18-RFC predominated among the three alternative clamp loaders, with mutants in Elg1-RFC or Rad24-RFC having less effect on trinucleotide repeats. Surprisingly, chl1, scc1-73, or scc2-4 mutants defective in sister chromatid cohesion (SCC) did not increase instability, suggesting that Ctf18-RFC protects triplet repeats independently of SCC. Instead, three results suggest novel roles for Ctf18-RFC in facilitating genomic stability. First, genetic instability in mutants of Ctf18-RFC was exacerbated by simultaneous deletion of the fork stabilizer Mrc1, but suppressed by deletion of the repair protein Rad52. Second, single-cell analysis showed that mutants in Ctf18-RFC had a slowed S phase and a striking G2/M accumulation, often with an abnormal multi-budded morphology. Third, ctf18 cells exhibit increased Rad52 foci in S phase, often persisting into G2, indicative of high levels of DNA damage. The presence of a repeat tract greatly magnified the ctf18 phenotypes. Together these results indicate that Ctf18-RFC has additional important functions in preserving genome stability, besides its role in SCC, which we propose include lesion bypass by replication forks and post-replication repair.
Vyšlo v časopise:
New Functions of Ctf18-RFC in Preserving Genome Stability outside Its Role in Sister Chromatid Cohesion. PLoS Genet 7(2): e32767. doi:10.1371/journal.pgen.1001298
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1001298
Souhrn
Expansion of DNA trinucleotide repeats causes at least 15 hereditary neurological diseases, and these repeats also undergo contraction and fragility. Current models to explain this genetic instability invoke erroneous DNA repair or aberrant replication. Here we show that CAG/CTG tracts are stabilized in Saccharomyces cerevisiae by the alternative clamp loader/unloader Ctf18-Dcc1-Ctf8-RFC complex (Ctf18-RFC). Mutants in Ctf18-RFC increased all three forms of triplet repeat instability—expansions, contractions, and fragility—with effect over a wide range of allele lengths from 20–155 repeats. Ctf18-RFC predominated among the three alternative clamp loaders, with mutants in Elg1-RFC or Rad24-RFC having less effect on trinucleotide repeats. Surprisingly, chl1, scc1-73, or scc2-4 mutants defective in sister chromatid cohesion (SCC) did not increase instability, suggesting that Ctf18-RFC protects triplet repeats independently of SCC. Instead, three results suggest novel roles for Ctf18-RFC in facilitating genomic stability. First, genetic instability in mutants of Ctf18-RFC was exacerbated by simultaneous deletion of the fork stabilizer Mrc1, but suppressed by deletion of the repair protein Rad52. Second, single-cell analysis showed that mutants in Ctf18-RFC had a slowed S phase and a striking G2/M accumulation, often with an abnormal multi-budded morphology. Third, ctf18 cells exhibit increased Rad52 foci in S phase, often persisting into G2, indicative of high levels of DNA damage. The presence of a repeat tract greatly magnified the ctf18 phenotypes. Together these results indicate that Ctf18-RFC has additional important functions in preserving genome stability, besides its role in SCC, which we propose include lesion bypass by replication forks and post-replication repair.
Zdroje
1. PearsonCE
EdamuraKN
ClearyJD
2005 Repeat instability: mechanisms of dynamic mutations. Nat Rev Genet 6 729 742
2. MirkinSM
2007 Expandable DNA repeats and human disease. Nature 447 932 940
3. FreudenreichCH
2007 Chromosome Fragility: Molecular mechanisms and cellular consequences. Frontiers in Bioscience 12 4911 4924
4. KovtunIV
McMurrayCT
2008 Features of trinucleotide repeat instability in vivo. Cell Res 18 198 213
5. LinY
DionV
WilsonJH
2006 Transcription promotes contraction of CAG repeat tracts in human cells. Nat Struct Mol Biol 13 179 180
6. YangZ
LauR
MarcadierJL
ChitayatD
PearsonCE
2003 Replication inhibitors modulate instability of an expanded trinucleotide repeat at the myotonic dystrophy type I disease locus in human cells. Am J Hum Genet 73 1092 1105
7. YoonS-R
DubeauL
de YoungM
WexlerNS
ArnheimN
2003 Huntington disease expansion mutations in humans can occur before meiosis is completed. Proc Natl Acad Sci USA 100 8834 8838
8. StrömL
KarlssonC
LindroosHB
WedahlS
KatouY
2007 Postreplicative formation of cohesion is required for repair and induced by a single DNA break. Science 317 242 245
9. ÜnalE
Heidinger-PauliJM
KoshlandD
2007 DNA double-strand breaks trigger genome-wide sister-chromatid cohesion through Eco1 (Ctf7). Science 317 245 248
10. SkibbensRV
2005 Unzipped and loaded: the role of DNA helicases and RFC clamp-loading complexes in sister chromatid cohesion. J Cell Biol 169 841 846
11. HannaJS
KrollES
LundbladV
SpencerFA
2001 Saccharomyces cerevisiae CTF18 and CTF4 are required for sister chromatid cohesion. Mol Cell Biol 21 3144 3158
12. MayerML
GygiSP
AebersoldR
HieterP
2001 Identification of RFC(Ctf18p, Ctf8p, Dcc1p): an alternative RFC complex required for sister chromatid cohesion in S. cerevisiae. Mol Cell 7 959 970
13. BermudezVP
ManiwaY
TappinI
OzatoK
HurwitzJ
2003 The alternative Ctf18-Dcc1-Ctf8-replication factor C complex required for sister chromatid cohesion loads proliferating cell nuclear antigen onto DNA. Proc Natl Acad Sci USA 100 10237 10242
14. ShiomiY
ShinozakiA
SugimotoK
UsukuraJ
ObuseC
2004 The reconstituted human Chl12-RFC complex functions as a second PCNA loader. Genes Cells 9 279 290
15. BylundGO
BurgersPMJ
2005 Replication protein A-directed unloading of PCNA by the Ctf18 cohesion establishment complex. Mol Cell Biol 25 5445 5455
16. LengronneA
McIntyreJ
KatouY
KanohY
HopfnerK-P
2006 Establishment of sister chromatid cohesion at the S. cerevisiae replication fork. Mol Cell 23 787 799
17. AnsbachAB
NoguchiC
KlansekIW
HeidlebaughM
NakamuraTM
2008 RFCCtf18 and the Swi1-Swi3 complex function in separate and redundant pathways required for the stabilization of replication forks to facilitate sister chromatid cohesion in Schizosaccharomyces pombe. Mol Biol Cell 19 595 607
18. CrabbeL
ThomasA
PantescoV
De VosJ
PaseroP
2010 Analysis of replication profiles reveals key role of RFC-Ctf18 in yeast replication stress response. Nat Struct Mol Biol 17 1391 1398
19. RazidloDF
LahueRS
2008 Mrc1, Tof1 and Csm3 inhibit CAG·CTG repeat instability by at least two mechanisms. DNA Repair 7 633 640
20. CallahanJL
AndrewsKJ
ZakianVA
FreudenreichCH
2003 Mutations in yeast replication proteins that increase CAG/CTG expansions also increase repeat fragility. Mol Cell Biol 23 7849 7860
21. CollinsSR
MillerKM
MaasNL
RoguevA
FillinghamJ
2007 Functional dissection of protein complexes involved in yeast chromosome biology using a genetic interaction map. Nature 446 806 810
22. XuH
BooneC
BrownGW
2007 Genetic dissection of parallel sister-chromatid cohesion pathways. Genetics 176 1417 1429
23. PetronczkiM
ChwallaB
SiomosMF
YokobayashiS
HelmhartW
2004 Sister-chromatid cohesion mediated by the alternative RFC-Ctf18/Dcc1/Ctf8, the helicase Chl1 and the polymerase-alpha-associated protein Ctf4 is essential for chromatid disjunction during meiosis II. J Cell Sci 117 3547 3559
24. SkibbensRV
2004 Chl1p, a DNA helicase-like protein in budding yeast, functions in sister-chromatid cohesion. Genetics 166 33 42
25. GambusA
van DeursenF
PolychronopoulosD
FoltmanM
JonesRC
2009 A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase α within the eukaryotic replisome. EMBO J 28 2992 3004
26. ErricoA
CosentinoC
RiveraT
LosadaA
SchwobE
2009 Tipin/Tim1/And1 protein complex promotes Polα chromatin binding and sister chromatid cohesion. EMBO J 28 3681 3692
27. SuterB
TongAHY
ChangM
YuL
BrownGW
2004 The origin recognition complex links replication, sister chromatid cohesion and transcriptional silencing in Saccharomyces cerevisiae. Genetics 167 579 591
28. KerrestA
AnandRP
SundararajanR
BermejoR
LiberiG
2009 SRS2 and SGS1 prevent chromosomal breaks and stabilize triplet repeats by restraining recombination. Nat Struct Mol Biol 16 159 167
29. SundararajanR
GellonL
ZunderRM
FreudenreichCH
2010 Double-strand break repair pathways protect against CAG/CTG repeat expansions, contractions and repeat-mediated chromosomal fragility in Saccharomyces cerevisiae. Genetics 184 65 77
30. GavinAC
AloyP
GrandiP
KrauseR
BoescheM
2006 Proteome survey reveals modularity of the yeast cell machinery. Nature 440 631 636
31. LouH
KomataM
KatouY
GuanZ
ReisC
2008 Mrc1 and DNA polymerase ε function together in linking DNA replication and the S phase checkpoint. Mol Cell 32 106 117
32. ZegermanP
DiffleyJFX
2003 Lessons in how to hold a fork. Nat Struct Biol 10 778
33. TourrièreH
PaseroP
2007 Maintenance of fork integrity at damaged DNA and natural pause sites. DNA Repair 6 900 913
34. LahiriM
GustafsonTL
MajorsER
FreudenreichCH
2004 Expanded CAG repeats activate the DNA damage checkpoint pathway. Mol Cell 15 287 293
35. FreudenreichCH
LahiriM
2004 Structure-forming CAG/CTG repeat sequences are sensitive to breakage in the absence of Mrc1 checkpoint function and S-phase checkpoint signaling: implications for trinucleotide repeat expansion diseases. Cell Cycle 3 1370 1374
36. PanX
YeP
YuanDS
WangX
BaderJS
2006 A DNA integrity network in the yeast Saccharomyces cerevisiae. Cell 124 1069 1081
37. EnserinkJM
SmolkaMB
ZhouH
KolodnerRD
2006 Checkpoint proteins control morphogenetic events during DNA replication stress in Saccharomyces cerevisiae. J Cell Biol 175 729 741
38. SandellLL
ZakianVA
1993 Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell 75 729 739
39. ToczyskiDP
HartwellLH
1997 CDC5 and CKII control adaptation to the yeast DNA damage checkpoint. Cell 90 1097 1106
40. LisbyM
BarlowJH
BurgessRC
RothsteinR
2004 Choreography of the DNA damage response: spatiotemporal relationships among checkpoint and repair proteins. Cell 118 699 713
41. SchmidtKH
KolodnerRD
2004 Requirement for Rrm3 helicase for repair of spontaneous DNA lesions in cells lacking Srs2 or Sgs1 helicase. Mol Cell Biol 24 3213 3226
42. MurakamiT
TakanoR
TakeoS
TaniguchiR
OgawaK
2010 Stable interaction between the human proliferating cell nuclear antigen loader complex Ctf18-replication factor C (RFC) and DNA polymerase epsilon is mediated by the cohesion-specific subunits, Ctf18, Dcc1, and Ctf8. J Biol Chem 285 34608 34615
43. KarrasGI
JentschS
2010 The RAD6 DNA damage tolerance pathway operates uncoupled from the replication fork and is functional beyond S phase. Cell 141 255 267
44. DaigakuY
DaviesAA
UlrichHD
2010 Ubiquitin-dependent DNA damage bypass is separable from genome replication. Nature 465 951 956
45. DaeeDL
MertzT
LahueRS
2007 Postreplication repair inhibits CAG·CTG repeat expansions in Saccharomyces cerevisiae. Mol Cell Biol 27 102 110
46. BranzeiD
VanoliF
FoianiM
2008 SUMOylation regulates Rad18-mediated template switch. Nature 456 915 920
47. TerretM-E
SherwoodR
RahmanS
QinJ
JallepalliPV
2009 Cohesin acetylation speeds the replication forks. Nature 462 231 234
48. OgiT
LimsirichaikulS
OvermeerRM
VolkerM
TakenakaK
2010 Three DNA polymerases, recruited by different mechanisms, carry out NER repair synthesis in human cells. Mol Cell 37 714 727
49. BrachmannCB
DaviesA
CostGJ
CaputoE
LiJ
1998 Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14 115 132
50. SlinkerBK
1998 The statistics of synergism. J Mol Cell Cardiol 30 723 731
51. ZhengQ
2002 Statistical and algorithmic methods for fluctuation analysis with SALVADOR as an implementation. Math Biosci 176 237 252
52. LisbyM
RothsteinR
MortensenUH
2001 Rad52 forms DNA repair and recombination centers during S phase. Proc Natl Acad Sci U S A 98 8276 8282
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2011 Číslo 2
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Meta-Analysis of Genome-Wide Association Studies in Celiac Disease and Rheumatoid Arthritis Identifies Fourteen Non-HLA Shared Loci
- MiRNA Control of Vegetative Phase Change in Trees
- The Cardiac Transcription Network Modulated by Gata4, Mef2a, Nkx2.5, Srf, Histone Modifications, and MicroRNAs
- Genome-Wide Transcript Profiling of Endosperm without Paternal Contribution Identifies Parent-of-Origin–Dependent Regulation of