Role of RecA and the SOS Response in Thymineless Death in
Thymineless death (TLD) is a classic and enigmatic phenomenon, documented in bacterial, yeast, and human cells, whereby cells lose viability rapidly when deprived of thymine. Despite its being the essential mode of action of important chemotherapeutic agents, and despite having been studied extensively for decades, the basic mechanisms of TLD have remained elusive. In Escherichia coli, several proteins involved in homologous recombination (HR) are required for TLD, however, surprisingly, RecA, the central HR protein and activator of the SOS DNA–damage response was reported not to be. We demonstrate that RecA and the SOS response are required for a substantial fraction of TLD. We show that some of the Rec proteins implicated previously promote TLD via facilitating activation of the SOS response and that, of the roughly 40 proteins upregulated by SOS, SulA, an SOS–inducible inhibitor of cell division, accounts for most or all of how SOS causes TLD. The data imply that much of TLD results from an irreversible cell-cycle checkpoint due to blocked cell division. FISH analyses of the DNA in cells undergoing TLD reveal blocked replication and apparent DNA loss with the region near the replication origin underrepresented initially and the region near the terminus lost later. Models implicating formation of single-strand DNA at blocked replication forks, a SulA-blocked cell cycle, and RecQ/RecJ-catalyzed DNA degradation and HR are discussed. The data predict the importance of DNA damage-response and HR networks to TLD and chemotherapy resistance in humans.
Vyšlo v časopise:
Role of RecA and the SOS Response in Thymineless Death in. PLoS Genet 6(3): e32767. doi:10.1371/journal.pgen.1000865
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1000865
Souhrn
Thymineless death (TLD) is a classic and enigmatic phenomenon, documented in bacterial, yeast, and human cells, whereby cells lose viability rapidly when deprived of thymine. Despite its being the essential mode of action of important chemotherapeutic agents, and despite having been studied extensively for decades, the basic mechanisms of TLD have remained elusive. In Escherichia coli, several proteins involved in homologous recombination (HR) are required for TLD, however, surprisingly, RecA, the central HR protein and activator of the SOS DNA–damage response was reported not to be. We demonstrate that RecA and the SOS response are required for a substantial fraction of TLD. We show that some of the Rec proteins implicated previously promote TLD via facilitating activation of the SOS response and that, of the roughly 40 proteins upregulated by SOS, SulA, an SOS–inducible inhibitor of cell division, accounts for most or all of how SOS causes TLD. The data imply that much of TLD results from an irreversible cell-cycle checkpoint due to blocked cell division. FISH analyses of the DNA in cells undergoing TLD reveal blocked replication and apparent DNA loss with the region near the replication origin underrepresented initially and the region near the terminus lost later. Models implicating formation of single-strand DNA at blocked replication forks, a SulA-blocked cell cycle, and RecQ/RecJ-catalyzed DNA degradation and HR are discussed. The data predict the importance of DNA damage-response and HR networks to TLD and chemotherapy resistance in humans.
Zdroje
1. AhmadSI
KirkSH
EisenstarkA
1998 Thymine metabolism and thymineless death in prokaryotes and eukaryotes. Annu Rev Microbiol 52 591 625
2. LongleyDB
HarkinDP
JohnstonPG
2003 5-fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 3 330 338
3. JackmanAL
TaylorGA
GibsonW
KimbellR
BrownM
1991 ICI D1694, a quinazoline antifolate thymidylate synthase inhibitor that is a potent inhibitor of L1210 tumor cell growth in vitro and in vivo: a new agent for clinical study. Cancer Res 51 5579 5586
4. SatB
RechesM
Engelberg-KulkaH
2003 The Escherichia coli mazEF suicide module mediates thymineless death. J Bacteriol 185 1803 1807
5. GodoyVG
JaroszDF
WalkerFL
SimmonsLA
WalkerGC
2006 Y-family DNA polymerases respond to DNA damage-independent inhibition of replication fork progression. Embo J 25 868 879
6. NakayamaH
HanawaltP
1975 Sedimentation analysis of deoxyribonucleic acid from thymine-starved Escherichia coli. J Bacteriol 121 537 547
7. CummingsDJ
KusyAR
1969 Thymineless death in Escherichia coli: inactivation and recovery. J Bacteriol 99 558 566
8. MorganrothPA
HanawaltPC
2006 Role of DNA replication and repair in thymineless death in Escherichia coli. J Bacteriol 188 5286 5288
9. AizenmanE
Engelberg-KulkaH
GlaserG
1996 An Escherichia coli chromosomal “addiction module” regulated by guanosine [corrected] 3′,5′-bispyrophosphate: a model for programmed bacterial cell death. Proc Natl Acad Sci U S A 93 6059 6063
10. NakayamaH
NakayamaK
NakayamaR
NakayamaY
1982 Recombination-deficient mutations and thymineless death in Escherichia coli K12: reciprocal effects of recBC and recF and indifference of recA mutations. Can J Microbiol 28 425 430
11. NakayamaK
ShiotaS
NakayamaH
1988 Thymineless death in Escherichia coli mutants deficient in the RecF recombination pathway. Can J Microbiol 34 905 907
12. NakayamaK
IrinoN
NakayamaH
1985 The recQ gene of Escherichia coli K12: molecular cloning and isolation of insertion mutants. Mol Gen Genet 200 266 271
13. NakayamaH
NakayamaK
NakayamaR
IrinoN
NakayamaY
1984 Isolation and genetic characterization of a thymineless death-resistant mutant of Escherichia coli K12: identification of a new mutation (recQ1) that blocks the RecF recombination pathway. Mol Gen Genet 195 474 480
14. SiegalE
1973 Ultraviolet-Sensitive Mutator Strain of Escherichia coli K-12. J Bacteriol 113 145 160
15. VeauteX
DelmasS
SelvaM
JeussetJ
Le CamE
2005 UvrD helicase, unlike Rep helicase, dismantles RecA nucleoprotein filaments in Escherichia coli. Embo J 24 180 189
16. FreifelderD
1969 Single-strand breaks in bacterial DNA associated with thymine starvation. J Mol Biol 45 1 7
17. BreitmanTR
MauryPB
ToalJN
1972 Loss of deoxyribonucleic acid-thymine during thymine starvation of Escherichia coli. J Bacteriol 112 646 648
18. NakayamaK
KusanoK
IrinoN
NakayamaH
1994 Thymine starvation-induced structural changes in Escherichia coli DNA. Detection by pulsed field gel electrophoresis and evidence for involvement of homologous recombination. J Mol Biol 243 611 620
19. AndersonJA
BarbourSD
1973 Effect of thymine starvation on deoxyribonucleic acid repair systems of Escherichia coli K-12. J Bacteriol 113 114 121
20. LauderSD
KowalczykowskiSC
1993 Negative co-dominant inhibition of recA protein function. Biochemical properties of the recA1, recA13 and recA56 proteins and the effect of recA56 protein on the activities of the wild-type recA protein function in vitro. J Mol Biol 234 72 86
21. MountDW
1971 Isolation and genetic analysis of a strain of Escherichia coli K-12 with an amber recA mutation. J Bacteriol 107 388 389
22. DutreixM
MoreauPL
BailoneA
GalibertF
BattistaJR
1989 New recA mutations that dissociate the various RecA protein activities in Escherichia coli provide evidence for an additional role for RecA protein in UV mutagenesis. J Bacteriol 171 2415 2423
23. InouyeM
1971 Pleiotropic effect of the recA gene of Escherichia coli: uncoupling of cell division from deoxyribonucleic acid replication. J Bacteriol 106 539 542
24. HuismanO
D'AriR
GeorgeJ
1980 Inducible sfi dependent division inhibition in Escherichia coli. Mol Gen Genet 177 629 636
25. HoweWE
MountDW
1975 Production of cells without deoxyribonucleic acid during thymidine starvation of lexA− cultures of Escherichia coli K-12. J Bacteriol 124 1113 1121
26. EnnisDG
FisherB
EdmistonS
MountDW
1985 Dual role for Escherichia coli RecA protein in SOS mutagenesis. Proc Natl Acad Sci U S A 82 3325 3329
27. Engelberg-KulkaH
HazanR
AmitaiS
2005 mazEF: a chromosomal toxin-antitoxin module that triggers programmed cell death in bacteria. J Cell Sci 118 4327 4332
28. LusettiSL
CoxMM
2002 The bacterial RecA protein and the recombinational DNA repair of stalled replication forks. Annu Rev Biochem 71 71 100
29. ButalaM
Zgur-BertokD
BusbySJ
2009 The bacterial LexA transcriptional repressor. Cell Mol Life Sci 66 82 93
30. FriedbergE
WalkerG
SiedeW
WoodR
SchultzR
2005 DNA Repair and Mutagenesis, Second Edition. Washington, D.C. ASM Press
31. MagnerDB
BlankschienMD
LeeJA
PenningtonJM
LupskiJR
2007 RecQ promotes toxic recombination in cells lacking recombination intermediate-removal proteins. Mol Cell 26 273 286
32. ZerbibD
MezardC
GeorgeH
WestSC
1998 Coordinated actions of RuvABC in Holliday junction processing. J Mol Biol 281 621 630
33. BoltEL
LloydRG
2002 Substrate specificity of RusA resolvase reveals the DNA structures targeted by RuvAB and RecG in vivo. Mol Cell 10 187 198
34. CourcelleJ
Carswell-CrumptonC
HanawaltPC
1997 recF and recR are required for the resumption of replication at DNA replication forks in Escherichia coli. Proc Natl Acad Sci U S A 94 3714 3719
35. RangarajanS
WoodgateR
GoodmanMF
2002 Replication restart in UV-irradiated Escherichia coli involving pols II, III, V, PriA, RecA and RecFOR proteins. Mol Microbiol 43 617 628
36. HishidaT
HanYW
ShibataT
KubotaY
IshinoY
2004 Role of the Escherichia coli RecQ DNA helicase in SOS signaling and genome stabilization at stalled replication forks. Genes Dev 18 1886 1897
37. CourcelleJ
HanawaltPC
1999 RecQ and RecJ process blocked replication forks prior to the resumption of replication in UV-irradiated Escherichia coli. Mol Gen Genet 262 543 551
38. SuskiC
MariansKJ
2008 Resolution of converging replication forks by RecQ and Topoisomerase III. Mol Cell 30 779 789
39. RaynardS
BussenW
SungP
2006 A double Holliday junction dissolvasome comprising BLM, Topoisomerase IIIalpha, and BLAP75. J Biol Chem 281 13861 13864
40. BatesD
KlecknerN
2005 Chromosome and replisome dynamics in E. coli: loss of sister cohesion triggers global chromosome movement and mediates chromosome segregation. Cell 121 899 911
41. PenningtonJM
RosenbergSM
2007 Spontaneous DNA breakage in single living Escherichia coli cells. Nat Genet 39 797 802
42. FonvilleNC
BlankschienMD
MagnerDB
RosenbergSM
2010 RecQ-dependent death-by-recombination in cells lacking RecG and UvrD. DNA Repair (in press) doi:10.1016/j.dnarep.2009.12.019
43. GuarinoE
SalgueroI
Jimenez-SanchezA
GuzmanEC
2007 Double-strand break generation under deoxyribonucleotide starvation in Escherichia coli. J Bacteriol 189 5782 5786
44. IpSC
BreguM
BarreFX
SherrattDJ
2003 Decatenation of DNA circles by FtsK-dependent Xer site-specific recombination. Embo J 22 6399 6407
45. PowellSN
KachnicLA
2008 Therapeutic exploitation of tumor cell defects in homologous recombination. Anticancer Agents Med Chem 8 448 460
46. SinghDK
AhnB
BohrVA
2009 Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10 235 252
47. MillerJ
1972 Generalized transduction; use of P1 in strain construction;
MillerJ
Cold Spring Harbor, New York Cold Spring Harbor Laboratory 201 205
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
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