rep

Summary

Gene Symbol: rep
Description: DNA helicase and single-stranded DNA-dependent ATPase
Alias: ECK3770, JW5604, dasC, mbrA, mmrA?
Species: Escherichia coli str. K-12 substr. MG1655

Top Publications

  1. Yarranton G, Gefter M. Enzyme-catalyzed DNA unwinding: studies on Escherichia coli rep protein. Proc Natl Acad Sci U S A. 1979;76:1658-62 pubmed
    ..form (RF) I DNA of bacteriophage varphiX174 requires the phage-induced cistron A (cisA) protein, the host rep protein, DNA-binding protein, ATP, and DNA polymerase III plus replication factors...
  2. Korolev S, Hsieh J, Gauss G, Lohman T, Waksman G. Major domain swiveling revealed by the crystal structures of complexes of E. coli Rep helicase bound to single-stranded DNA and ADP. Cell. 1997;90:635-47 pubmed
    Crystal structures of binary and ternary complexes of the E. coli Rep helicase bound to single-stranded (ss) DNA or ssDNA and ADP were determined to a resolution of 3.0 A and 3.2 A, respectively...
  3. Lestini R, Michel B. UvrD and UvrD252 counteract RecQ, RecJ, and RecFOR in a rep mutant of Escherichia coli. J Bacteriol. 2008;190:5995-6001 pubmed publisher
    b>Rep and UvrD are two related Escherichia coli helicases, and inactivating both is lethal...
  4. Myong S, Rasnik I, Joo C, Lohman T, Ha T. Repetitive shuttling of a motor protein on DNA. Nature. 2005;437:1321-5 pubmed
    ..developed a single-molecule fluorescence assay that can visualize DNA binding and translocation of Escherichia coli Rep, a superfamily 1 DNA helicase homologous to Saccharomyces cerevisiae Srs2...
  5. Sandler S. Multiple genetic pathways for restarting DNA replication forks in Escherichia coli K-12. Genetics. 2000;155:487-97 pubmed
    ..different priA2::kan suppressors (both dnaC alleles) were tested for their ability to rescue the priA-priC and priA-rep double mutant lethality. Only dnaC809,820 (and not dnaC809) could rescue the lethality in each case...
  6. Brendza K, Cheng W, Fischer C, Chesnik M, Niedziela Majka A, Lohman T. Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain. Proc Natl Acad Sci U S A. 2005;102:10076-81 pubmed
    ..The Escherichia coli Rep protein, a superfamily 1 DNA helicase, functions in DNA replication restart and is required for replication of ..
  7. Heller R, Marians K. Non-replicative helicases at the replication fork. DNA Repair (Amst). 2007;6:945-52 pubmed
    ..Here we examine the DNA-binding specificity of three of the latter group, PriA, Rep, and UvrD. Only PriA and Rep display structure-specific fork binding...
  8. Lane H, Denhardt D. The rep mutation. IV. Slower movement of replication forks in Escherichia coli rep strains. J Mol Biol. 1975;97:99-112 pubmed
  9. Cheng W, Hsieh J, Brendza K, Lohman T. E. coli Rep oligomers are required to initiate DNA unwinding in vitro. J Mol Biol. 2001;310:327-50 pubmed
    E. coli Rep protein is a 3' to 5' SF1 superfamily DNA helicase which is monomeric in the absence of DNA, but can dimerize upon binding either single-stranded or duplex DNA...

More Information

Publications68

  1. Ha T, Rasnik I, Cheng W, Babcock H, Gauss G, Lohman T, et al. Initiation and re-initiation of DNA unwinding by the Escherichia coli Rep helicase. Nature. 2002;419:638-41 pubmed
    ..assays--which do not interfere with biological activity--to probe DNA unwinding by the Escherichia coli Rep helicase...
  2. Veaute X, Delmas S, Selva M, Jeusset J, Le Cam E, Matic I, et al. UvrD helicase, unlike Rep helicase, dismantles RecA nucleoprotein filaments in Escherichia coli. EMBO J. 2005;24:180-9 pubmed
    The roles of UvrD and Rep DNA helicases of Escherichia coli are not yet fully understood. In particular, the reason for rep uvrD double mutant lethality remains obscure...
  3. Trun N, Gottesman S, Løbner Olesen A. Analysis of Escherichia coli mutants with altered DNA content. Cold Spring Harb Symp Quant Biol. 1991;56:353-8 pubmed
  4. Sevastsyanovich Y, Titok M, Krasowiak R, Bingle L, Thomas C. Ability of IncP-9 plasmid pM3 to replicate in Escherichia coli is dependent on both rep and par functions. Mol Microbiol. 2005;57:819-33 pubmed
    ..A 1.3 kb ori V-rep fragment from IncP-9 plasmid pM3 was sufficient for autonomous replication in Pseudomonas putida but not in ..
  5. Arai N, Arai K, Kornberg A. Complexes of Rep protein with ATP and DNA as a basis for helicase action. J Biol Chem. 1981;256:5287-93 pubmed
    The rep protein of Escherichia coli, a helicase that unwinds duplex DNA at a replication fork (Kornberg, A., Scott, J. F., and Bertsch, L. L. (1978) J. Biol. Chem...
  6. Heller R, Marians K. The disposition of nascent strands at stalled replication forks dictates the pathway of replisome loading during restart. Mol Cell. 2005;17:733-43 pubmed
    ..These observations suggest that the type of initial damage on the DNA template and how the inactivated fork is processed ultimately influence the choice of enzymatic restart pathway. ..
  7. Yancey J, Matson S. The DNA unwinding reaction catalyzed by Rep protein is facilitated by an RHSP-DNA interaction. Nucleic Acids Res. 1991;19:3943-51 pubmed
    The unwinding reaction catalyzed by the Escherichia coli Rep protein is stimulated by a small 15 kDa protein called Rep helicase stimulatory protein (RHSP)(1)...
  8. Bialkowska Hobrzanska H, Gilchrist C, Denhardt D. Escherichia coli rep gene: identification of the promoter and N terminus of the rep protein. J Bacteriol. 1985;164:1004-10 pubmed
    The functional Escherichia coli rep gene, which encodes the Mr 67,000 Rep helicase, has been localized within a 2.55-kilobase sequence. Its regulatory region has been characterized by the use of rep-lacZ fusions...
  9. Trun N, Gottesman S. On the bacterial cell cycle: Escherichia coli mutants with altered ploidy. Genes Dev. 1990;4:2036-47 pubmed
    ..b>mbrA maps to 68 min on the E. coli chromosome, mbrB to 88.5 min, mbrC to 89.5 min, and mbrD to 90 min...
  10. Lohman T. Escherichia coli DNA helicases: mechanisms of DNA unwinding. Mol Microbiol. 1992;6:5-14 pubmed
    ..coli Rep, RecBCD and phage T7 gene 4 helicases.
  11. Shiraishi K, Imai Y, Yoshizaki S, Ikeda H. Rep helicase suppresses short-homology-dependent illegitimate recombination in Escherichia coli. Genes Cells. 2005;10:1015-23 pubmed
    To study roles of Rep helicase in short-homology-dependent illegitimate recombination, we examined the effect of a rep mutation on illegitimate recombination and found that the frequency of spontaneous illegitimate recombination is ..
  12. Moore K, Lohman T. Kinetic mechanism of adenine nucleotide binding to and hydrolysis by the Escherichia coli Rep monomer. 1. Use of fluorescent nucleotide analogues. Biochemistry. 1994;33:14550-64 pubmed
    The Escherichia coli Rep helicase catalyzes the unwinding of duplex DNA in a reaction that is coupled to ATP binding and hydrolysis...
  13. Guy C, Atkinson J, Gupta M, Mahdi A, Gwynn E, Rudolph C, et al. Rep provides a second motor at the replisome to promote duplication of protein-bound DNA. Mol Cell. 2009;36:654-66 pubmed publisher
    ..We show that Rep and UvrD each promote movement of E...
  14. Maisnier Patin S, Nordstrom K, DasGupta S. Replication arrests during a single round of replication of the Escherichia coli chromosome in the absence of DnaC activity. Mol Microbiol. 2001;42:1371-82 pubmed
    ..In a dnaC2(Ts)rep double mutant, almost all cells failed to complete chromosome replication in the absence of DnaC activity...
  15. Sladewski T, Hetrick K, Foster P. Escherichia coli Rep DNA helicase and error-prone DNA polymerase IV interact physically and functionally. Mol Microbiol. 2011;80:524-41 pubmed publisher
    ..Here we demonstrate that Pol IV interacts in vitro with Rep DNA helicase and that this interaction enhances Rep's helicase activity...
  16. Okumus B, Arslan S, Fengler S, Myong S, Ha T. Single molecule nanocontainers made porous using a bacterial toxin. J Am Chem Soc. 2009;131:14844-9 pubmed publisher
    ..The DNA translocation activity of E. coli Rep helicase inside vesicles was fueled by ATP provided outside the vesicle, and a dramatically higher number of ..
  17. Tessman I, Fassler J, Bennett D. Relative map location of the rep and rho genes of Escherichia coli. J Bacteriol. 1982;151:1637-40 pubmed
    The rep gene of Escherichia coli was mapped between ilvC and rho by three-factor P1 transductional crosses and also by complementation with a set of lambda transducing phages that contain known amounts of bacterial DNA linked to ilvC...
  18. Jankowsky E. Biophysics: helicase snaps back. Nature. 2005;437:1245 pubmed
  19. Wong I, Lohman T. ATPase activity of Escherichia coli Rep helicase crosslinked to single-stranded DNA: implications for ATP driven helicase translocation. Proc Natl Acad Sci U S A. 1996;93:10051-6 pubmed
    ..to helicase translocation along DNA, we have purified and characterized complexes of the Escherichia coli Rep protein, a dimeric DNA helicase, covalently crosslinked to a single-stranded hexadecameric oligodeoxynucleotide (S)...
  20. Cheng W, Brendza K, Gauss G, Korolev S, Waksman G, Lohman T. The 2B domain of the Escherichia coli Rep protein is not required for DNA helicase activity. Proc Natl Acad Sci U S A. 2002;99:16006-11 pubmed
    The Escherichia coli Rep protein is a 3' to 5' SF1 DNA helicase required for replication of bacteriophage phiX174 in E. coli, and is structurally homologous to the E. coli UvrD helicase and the Bacillus stearothermophilus PcrA helicase...
  21. Wong I, Amaratunga M, Lohman T. Heterodimer formation between Escherichia coli Rep and UvrD proteins. J Biol Chem. 1993;268:20386-91 pubmed
    ..although Escherichia coli can survive mutations or deletions of either the uvrD gene (encoding Helicase II) or the rep gene (encoding Rep helicase) separately, deletion of both rep and uvrD genes is lethal (Washburn, B. K...
  22. Amaratunga M, Lohman T. Escherichia coli rep helicase unwinds DNA by an active mechanism. Biochemistry. 1993;32:6815-20 pubmed
    ..We have examined the kinetics of DNA unwinding by Escherichia coli Rep protein (a 3' to 5' helicase) by rapid quench-flow methods using a series of novel, nonnatural DNA substrates ..
  23. Gilchrist C, Denhardt D. Escherichia coli rep gene: sequence of the gene, the encoded helicase, and its homology with uvrD. Nucleic Acids Res. 1987;15:465-75 pubmed
    The sequence of a 2.67-kilobase section of the Escherichia coli chromosome that contains the rep gene has been determined...
  24. Lohman T, Bjornson K. Mechanisms of helicase-catalyzed DNA unwinding. Annu Rev Biochem. 1996;65:169-214 pubmed
    ..Recent studies of DNA and NTP binding and DNA unwinding by the Escherichia coli (E. coli) Rep helicase suggest that the Rep helicase dimer unwinds DNA by an active, rolling mechanism...
  25. Baharoglu Z, Petranovic M, Flores M, Michel B. RuvAB is essential for replication forks reversal in certain replication mutants. EMBO J. 2006;25:596-604 pubmed
    ..mutants dnaEts and holD, affected for the Pol III catalytic subunit and clamp loader, and in the helicase mutant rep. This finding reveals that the recombination enzyme RuvAB targets forks in vivo and we propose that it directly ..
  26. Arai N, Kornberg A. Rep protein as a helicase in an active, isolatable replication fork of duplex phi X174 DNA. J Biol Chem. 1981;256:5294-8 pubmed
    b>Rep protein as a helicase combines its actions with those of gene A protein and single-stranded DNA binding protein to separate the strands of phi X174 duplex DNA and thereby can generate and advance a replication fork (Scott, J. F...
  27. Smith K, Yancey J, Matson S. Identification and purification of a protein that stimulates the helicase activity of the Escherichia coli Rep protein. J Biol Chem. 1989;264:6119-26 pubmed
    ..coli Rep protein...
  28. Aldea M, Maples V, Kushner S. Generation of a detailed physical and genetic map of the ilv-metE-udp region of the Escherichia coli chromosome. J Mol Biol. 1988;200:427-38 pubmed
  29. Driver R, Lawther R. Physical analysis of deletion mutations in the ilvGEDA operon of Escherichia coli K-12. J Bacteriol. 1985;162:598-606 pubmed
    ..base pairs from within ilvD, whereas the largest mutation resulted from the deletion of 17 kilobases including the rep gene...
  30. Harrington E, Trun N. Unfolding of the bacterial nucleoid both in vivo and in vitro as a result of exposure to camphor. J Bacteriol. 1997;179:2435-9 pubmed
    ..We show that exposure of isolated nucleoids to camphor results in unfolding of the chromosome. ..
  31. Takahashi S, Hours C, Chu A, Denhardt D. The rep mutation. VI. Purification and properties of the Escherichia coli rep protein, DNA helicase III. Can J Biochem. 1979;57:855-66 pubmed
    The protein product of the rep gene of Escherichia coli is required for the replication of certain bacteriophage genomes (phi X174, fd, P2) and for the normal replication of E. coli DNA...
  32. Thompson K, Rhodius V, Gottesman S. SigmaE regulates and is regulated by a small RNA in Escherichia coli. J Bacteriol. 2007;189:4243-56 pubmed
    ..A number of plasmids, including some carrying rybB, negatively regulated the fusion. An insertion in the rep helicase and one upstream of dnaK decreased expression of the fusion...
  33. Sandler S. Requirements for replication restart proteins during constitutive stable DNA replication in Escherichia coli K-12. Genetics. 2005;169:1799-806 pubmed
    ..It is also shown that rep and rnhA mutations are synthetically lethal. DnaC809 and dnaC809,820 mutations suppress this lethality...
  34. Boonsombat R, Yeh S, Milne A, Sandler S. A novel dnaC mutation that suppresses priB rep mutant phenotypes in Escherichia coli K-12. Mol Microbiol. 2006;60:973-83 pubmed
    ..During replication restart in Escherichia coli, the PriA, PriB, PriC, DnaT and Rep proteins form multiple pathways that bind to repaired replication forks...
  35. Lane H, Denhardt D. The rep mutation. III. Altered structure of the replicating Escherichia coli chromosome. J Bacteriol. 1974;120:805-14 pubmed
    The rep gene function of Escherichia coli is essential for the replication of P2 and phiX174 double-stranded deoxyribonucleic acid (DNA)...
  36. Calendar R, Lindqvist B, Sironi G, Clark A. Characterization of REP- mutants and their interaction with P2 phage. Virology. 1970;40:72-83 pubmed
  37. Michel B, Recchia G, Penel Colin M, Ehrlich S, Sherratt D. Resolution of holliday junctions by RuvABC prevents dimer formation in rep mutants and UV-irradiated cells. Mol Microbiol. 2000;37:180-91 pubmed
    ..First, although arrested replication forks are rescued by recombinational repair in cells deficient for the Rep helicase, rep mutants do not require the XerCD proteins or the dif site for viability...
  38. Wong I, Lohman T. Allosteric effects of nucleotide cofactors on Escherichia coli Rep helicase-DNA binding. Science. 1992;256:350-5 pubmed
    The Escherichia coli Rep helicase unwinds duplex DNA during replication. The functional helicase appears to be a dimer that forms only on binding DNA. Both protomers of the dimer can bind either single-stranded or duplex DNA...
  39. Baylor N, Williams A, Cofie N. Molecular characterization of ilvC specialized transducing phages of Escherichia coli K-12. Mol Gen Genet. 1983;191:347-52 pubmed
    ..have shown that the lambda defective ilvC phages were able to cotransduce a rho-15ts mutation as well as a rep-5 mutation. Hence, the order of the clockwise excision of the ilv cluster was found to be ilvC-rho-rep-cya...
  40. Michel B, Ehrlich S, Uzest M. DNA double-strand breaks caused by replication arrest. EMBO J. 1997;16:430-8 pubmed
    ..Two E. coli replicative helicases were tested, Rep, which is essential in recBC mutants, and DnaB...
  41. Trun N. Mutations in the E. coli Rep helicase increase the amount of DNA per cell. FEMS Microbiol Lett. 2003;226:187-93 pubmed
    ..Eight independently isolated alleles of rep(mbrA) contain the same three changes in the rep gene: a G to A at position -44 from the start of the mRNA (+1); an opal ..
  42. Yancey Wrona J, Wood E, George J, Smith K, Matson S. Escherichia coli Rep protein and helicase IV. Distributive single-stranded DNA-dependent ATPases that catalyze a limited unwinding reaction in vitro. Eur J Biochem. 1992;207:479-85 pubmed
    b>Rep protein and helicase IV, two DNA-dependent adenosine 5'-triphosphatases with helicase activity, have been purified from Escherichia coli and characterized...
  43. Trun N, Gottesman S. Characterization of Escherichia coli mutants with altered ploidy. Res Microbiol. 1991;142:195-200 pubmed
    ..The mutants (called mbr for moth-ball-resistant) map to four chromosomal locations: mbrA at 68 min; mbrB at 88.5 min; mbrC at 89.5 min; and mbrD at 90 min...
  44. Bjornson K, Wong I, Lohman T. ATP hydrolysis stimulates binding and release of single stranded DNA from alternating subunits of the dimeric E. coli Rep helicase: implications for ATP-driven helicase translocation. J Mol Biol. 1996;263:411-22 pubmed
    ..To probe the mechanism of ATP-driven translocation by the dimeric E. coli Rep helicase along single stranded (ss) DNA, we examined the effects of ATP on the dissociation kinetics of ssDNA from ..
  45. Boubakri H, de Septenville A, Viguera E, Michel B. The helicases DinG, Rep and UvrD cooperate to promote replication across transcription units in vivo. EMBO J. 2010;29:145-57 pubmed publisher
    ..We report here that in vivo, the DinG, Rep and UvrD helicases are essential for efficient replication across highly transcribed regions...
  46. Wong I, Chao K, Bujalowski W, Lohman T. DNA-induced dimerization of the Escherichia coli rep helicase. Allosteric effects of single-stranded and duplex DNA. J Biol Chem. 1992;267:7596-610 pubmed
    The Escherichia coli Rep helicase is a stable monomer (Mr = 72,802) in the absence of DNA; however, binding of single-stranded (ss) or duplex (ds) DNA induces Rep monomers to dimerize...
  47. Bjornson K, Hsieh J, Amaratunga M, Lohman T. Kinetic mechanism for the sequential binding of two single-stranded oligodeoxynucleotides to the Escherichia coli Rep helicase dimer. Biochemistry. 1998;37:891-9 pubmed
    Escherichia coli Rep helicase is a DNA motor protein that unwinds duplex DNA as a dimeric enzyme...
  48. Rasnik I, Myong S, Cheng W, Lohman T, Ha T. DNA-binding orientation and domain conformation of the E. coli rep helicase monomer bound to a partial duplex junction: single-molecule studies of fluorescently labeled enzymes. J Mol Biol. 2004;336:395-408 pubmed
    ..Crystal structures of two such helicases, Escherichia coli Rep and Bacillus stearothermophilus PcrA, show that the 2B sub-domain of these proteins can be found in dramatically ..
  49. Lohman T, Chao K, Green J, Sage S, Runyon G. Large-scale purification and characterization of the Escherichia coli rep gene product. J Biol Chem. 1989;264:10139-47 pubmed
    We report a procedure for the large-scale purification of the Escherichia coli Rep protein, a helicase that is involved in the replication of the E. coli chromosome as well as a number of single-stranded bacteriophages...
  50. Wong I, Moore K, Bjornson K, Hsieh J, Lohman T. ATPase activity of Escherichia coli Rep helicase is dramatically dependent on DNA ligation and protein oligomeric states. Biochemistry. 1996;35:5726-34 pubmed
    The Escherichia coli Rep helicase catalyzes the unwinding of duplex DNA using the energy derived from ATP binding and hydrolysis. Rep functions as a dimer but assembles to its active dimeric form only on binding DNA...
  51. Bjornson K, Moore K, Lohman T. Kinetic mechanism of DNA binding and DNA-induced dimerization of the Escherichia coli Rep helicase. Biochemistry. 1996;35:2268-82 pubmed
    The monomeric Escherichia coli Rep protein undergoes a DNA-induced dimerization upon binding either single-stranded (ss) or duplex DNA with the dimer being the active form of the Rep helicase...
  52. Bjornson K, Amaratunga M, Moore K, Lohman T. Single-turnover kinetics of helicase-catalyzed DNA unwinding monitored continuously by fluorescence energy transfer. Biochemistry. 1994;33:14306-16 pubmed
    ..substrate in pre-steady state, single turnover studies of duplex DNA unwinding catalyzed by the Escherichia coli Rep helicase, monitored by fluorescence stopped flow...
  53. Moore K, Lohman T. Kinetic mechanism of adenine nucleotide binding to and hydrolysis by the Escherichia coli Rep monomer. 2. Application of a kinetic competition approach. Biochemistry. 1994;33:14565-78 pubmed
    The Escherichia coli Rep protein is a DNA helicase that functions as a homodimer to catalyze the unwinding of duplex DNA during DNA replication in a reaction that is coupled to the binding and hydrolysis of ATP...
  54. Hsieh J, Moore K, Lohman T. A two-site kinetic mechanism for ATP binding and hydrolysis by E. coli Rep helicase dimer bound to a single-stranded oligodeoxynucleotide. J Mol Biol. 1999;288:255-74 pubmed
    Escherichia coli Rep helicase catalyzes the unwinding of duplex DNA in reactions that are coupled to ATP binding and hydrolysis...
  55. Colasanti J, Denhardt D. The Escherichia coli rep mutation. X. Consequences of increased and decreased Rep protein levels. Mol Gen Genet. 1987;209:382-90 pubmed
    Recombinant DNA techniques were used to study various aspects of rep gene function in Escherichia coli...
  56. Chao K, Lohman T. DNA-induced dimerization of the Escherichia coli Rep helicase. J Mol Biol. 1991;221:1165-81 pubmed
    The Escherichia coli Rep protein is a DNA helicase that is involved in DNA replication...
  57. Chao K, Lohman T. DNA and nucleotide-induced conformational changes in the Escherichia coli Rep and helicase II (UvrD) proteins. J Biol Chem. 1990;265:1067-76 pubmed
    The domain structures of the Escherichia coli Rep and Helicase II proteins and their ligand-dependent conformational changes have been examined by monitoring the sensitivity of these helicases to proteolysis by trypsin and chymotrypsin...
  58. Białkowska Hobrzanska H, Denhardt D. The rep mutation. VII. Cloning and analysis of the functional rep gene of Escherichia coli K-12. Gene. 1984;28:93-102 pubmed
    The rep gene of Escherichia coli was isolated on a 6-kb PvuII fragment of plasmid pLC44-7 DNA from the Clarke-Carbon collection and cloned into pSC101 (to form pHBH8) and pBR322 (to form pHBH30)...
  59. Bentchikou E, Chagneau C, Long E, Matelot M, Allemand J, Michel B. Are the SSB-Interacting Proteins RecO, RecG, PriA and the DnaB-Interacting Protein Rep Bound to Progressing Replication Forks in Escherichia coli?. PLoS ONE. 2015;10:e0134892 pubmed publisher
    ..Neither the recombination proteins RecO and RecG, nor the replication accessory helicase Rep are detected specifically in replicating cells in our assay, indicating that either they are not present at ..