Genetic and molecular analyses of the C-terminal region of the recE gene from the Rac prophage of Escherichia coli K-12 reveal the recT gene
10.1128/jb.175.23.7673-7682.1993
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The nucleotide sequence of the C-terminal region of the recE gene of the Rac prophage of Escherichia coli K-12 reveals the presence of a partially overlapping reading frame we call recT. Deletion mutations show that recT is required for the RecE pathway of conjugational recombination. By cloning recT with a plasmid vector compatible with pBR322, we showed by cis-trans tests that the portion of the recE gene encoding ExoVIII DNA nuclease activity is also required for RecE pathway conjugational recombination. The recT gene can replace the redB gene of lambda for recA-independent plasmid recombination. A Tn10 insertion mutation previously thought to be in recE is located in recT and is renamed recT101::Tn10. Discrepancies between the molecular mass estimates of wild-type ExoVIII protein determined from mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and calculated from the predicted amino acid sequence are discussed. The hypothesis that wild-type ExoVIII protein results from fusion of RecE and RecT proteins is disproved genetically, thus supporting a previous hypothesis that the discrepancies are due to abnormal protein mobility in SDS-PAGE. A computer-performed scan of the bacteriophage nucleotide sequence data base of GenBank revealed substantial similarity between most of recE and a 2.5-kb portion of the b2 region of lambda. This suggests interesting speculations concerning the evolutionary relationship of lambda and Rac prophages.

Bibliography

Clark, A. J., Sharma, V., Brenowitz, S., Chu, C. C., Sandler, S., Satin, L., Templin, A., Berger, I., & Cohen, A. (1993). Genetic and molecular analyses of the C-terminal region of the recE gene from the Rac prophage of Escherichia coli K-12 reveal the recT gene. Journal of Bacteriology, 175(23), 7673–7682.

Authors 9
  1. A J Clark (first)
  2. V Sharma (additional)
  3. S Brenowitz (additional)
  4. C C Chu (additional)
  5. S Sandler (additional)
  6. L Satin (additional)
  7. A Templin (additional)
  8. I Berger (additional)
  9. A Cohen (additional)
References 40 Referenced 38
  1. Adelberg , E. A. , and S. M. Burns . 1960 . Genetic variation in the sex factor of Escherichia coli . J. Bacteriol. 79 : 321 - 330 . (10.1128/jb.79.3.321-330.1960) / J. Bacteriol. / Genetic variation in the sex factor of Escherichia coli by Adelberg E. A. (1960)
  2. Barbour , S. D. , H. Nagaishi , A. Templin , and A. J. Clarl . 1970 . Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants due to indirect suppression of Rec- mutations . Proc. Natl. Acad. Sci. USA 67 : 128 - 135 . (10.1073/pnas.67.1.128) / Proc. Natl. Acad. Sci. USA / Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants due to indirect suppression of Rec- mutations by Barbour S. D. (1970)
  3. Berger , I. , and A. Cohen . 1989 . Suppression of RecA deficiency in plasmid recombination by bacteriophage X ,Bprotein in RecBCD - Exol- Escherichia coli cells . J. Bacteriol. 171 : 3523 - 3529 . (10.1128/jb.171.6.3523-3529.1989) / J. Bacteriol. / Suppression of RecA deficiency in plasmid recombination by bacteriophage X ,Bprotein in RecBCD - Exol- Escherichia coli cells by Berger I. (1989)
  4. Birnboim , H. C. , and J. Doly . 1979 . A rapid alkaline extraction procedure for screening recombinant plasmid DNA . Nucleic Acids Res. 7 : 1513 - 1523 . (10.1093/nar/7.6.1513) / Nucleic Acids Res. / A rapid alkaline extraction procedure for screening recombinant plasmid DNA by Birnboim H. C. (1979)
  5. Brown , M. , A. Weston , J. Saunders , and G. Humphreys . 1979 . Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth . FEMS Microbiol. Lett. 5 : 219 - 222 . (10.1111/j.1574-6968.1979.tb03283.x) / FEMS Microbiol. Lett. / Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth by Brown M. (1979)
  6. Calos M. Personal communication.
  7. Calos , M. P. , J. S. Lebkowski , and M. R. Botchan . 1983 . High mutation frequency in DNA transfected into mammalian cells . Proc. Natl. Acad. Sci. USA 80 : 3015 - 3019 . (10.1073/pnas.80.10.3015) / Proc. Natl. Acad. Sci. USA / High mutation frequency in DNA transfected into mammalian cells by Calos M. P. (1983)
  8. Chang , A. C. Y. , and S. Cohen . 1978 . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from P1SA cryptic miniplasmid . J. Bacteriol. 134 : 1141 - 1156 . (10.1128/jb.134.3.1141-1156.1978) / J. Bacteriol. / Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from P1SA cryptic miniplasmid by Chang A. C. Y. (1978)
  9. Chen , E. Y. , and P. H. Seeberg . 1985 . Supercoil sequencing: a fast and simple method for sequencing plasmid DNA . DNA 4 : 165 - 170 . (10.1089/dna.1985.4.165) / DNA / Supercoil sequencing: a fast and simple method for sequencing plasmid DNA by Chen E. Y. (1985)
  10. Chu , C. C. , A. Templin , and A. J. Clark . 1989 . Suppression of a frameshift mutation in the recE gene of Escherichia coli K-12 occurs by gene fusion . J. Bacteriol. 171 : 2101 - 2109 . (10.1128/jb.171.4.2101-2109.1989) / J. Bacteriol. / Suppression of a frameshift mutation in the recE gene of Escherichia coli K-12 occurs by gene fusion by Chu C. C. (1989)
  11. Clark A. J. Unpublished data.
  12. Clark , A. J. 1963 . Genetic analysis of a 'double male' strain of Escherichia coli K-12 . Genetics 48 : 105 - 120 . (10.1093/genetics/48.1.105) / Genetics / Genetic analysis of a 'double male' strain of Escherichia coli K-12 by Clark A. J. (1963)
  13. Clark , A. J. , and A. D. Margulies . 1965 . Isolation and characterization of recombination-deficient mutants of Escherichia coli K-12 . Proc. Natl. Acad. Sci. USA 53 : 451 - 459 . (10.1073/pnas.53.2.451) / Proc. Natl. Acad. Sci. USA / Isolation and characterization of recombination-deficient mutants of Escherichia coli K-12 by Clark A. J. (1965)
  14. Clark , A. J. , S. J. Sandler , D. K. Willis , C. C. Chu , M. A. Blanar , and S. T. Lovett . 1984 . Genes of the RecE and RecF pathways of conjugational recombination in E. coli . Cold Spring Harbor Symp. Quant. Biol. 49 : 453 - 462 . (10.1101/SQB.1984.049.01.051) / Cold Spring Harbor Symp. Quant. Biol. / Genes of the RecE and RecF pathways of conjugational recombination in E. coli by Clark A. J. (1984)
  15. Daniels D. L. J. L. Schroeder W. Szybalski F. Sanger and F. R. Blattner. 1983. Appendix I: a molecular map of coliphage lambda p. 469-517. In R. W. Hendrix J. W. Roberts F. W. Stahl and R. A. Weisberg (ed.) Lambda II. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  16. Daniels D. L. J. L. Schroeder W. Szybalski F. Sanger A. R. Coulson G. R. Hong D. F. Hill G. B. Petersen and F. R. Blattner. 1983. Appendix II: complete annotated lambda sequence p. 529-676. In R. W. Hendrix J. W. Roberts F. W. Stahl and R. A. Weisberg (ed.) Lambda II. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  17. Demerec , M. , E. A. Adelberg , A. J. Clark , and P. E. Hartman . 1966 . A proposal for a uniform nomenclature in bacterial genetics . Genetics 54 : 61 - 76 . (10.1093/genetics/54.1.61) / Genetics / A proposal for a uniform nomenclature in bacterial genetics by Demerec M. (1966)
  18. Fouts , K. E. , T. Wasie-Gilbert , D. K. Willis , A. J. Clark , and S. D. Barbour . 1983 . Genetic analysis of transposon-induced mutations of the Rac prophage in Escherichia coli K-12 which affect expression of recE . J. Bacteriol. 156 : 718 - 726 . (10.1128/jb.156.2.718-726.1983) / J. Bacteriol. / Genetic analysis of transposon-induced mutations of the Rac prophage in Escherichia coli K-12 which affect expression of recE by Fouts K. E. (1983)
  19. Gillen , J. R. , A. E. Karu , H. Nagaishi , and A. J. Clark 1977 . Characterization of the deoxyribonuclease determined by lambda reverse as exonuclease VIII of Escherichia coli . J. Mol. Biol. 113 : 27 - 41 . (10.1016/0022-2836(77)90039-0) / J. Mol. Biol. / Characterization of the deoxyribonuclease determined by lambda reverse as exonuclease VIII of Escherichia coli by Gillen J. R. (1977)
  20. Hall , S. D. , M. F. Kane , and R. D. Kolodner . 1993 . Identification and characterization of the Escherichia coli RecT protein, a protein encoded by the recE region that promotes renaturation of homologous single-stranded DNA . J. Bacteriol. 175 : 277 - 287 . (10.1128/jb.175.1.277-287.1993) / J. Bacteriol. / Identification and characterization of the Escherichia coli RecT protein, a protein encoded by the recE region that promotes renaturation of homologous single-stranded DNA by Hall S. D. (1993)
  21. Halling , S. M. , R. W. Simons , J. C. Way , R. B. Walsh , and N. Kleckner . 1982 . DNA sequence organisation of IS10-right of TnlO and comparison with IS10-left . Proc. Natl. Acad. Sci. USA 79 : 2608 - 2612 . (10.1073/pnas.79.8.2608) / Proc. Natl. Acad. Sci. USA / DNA sequence organisation of IS10-right of TnlO and comparison with IS10-left by Halling S. M. (1982)
  22. Henikoff , S. 1984 . Unidirectional digestion with exonuclease III creates targeted breakpoint for DNA sequencing . Gene 28 : 351 - 359 . (10.1016/0378-1119(84)90153-7) / Gene / Unidirectional digestion with exonuclease III creates targeted breakpoint for DNA sequencing by Henikoff S. (1984)
  23. Joseph , J. W. , and R. Kolodner . 1983 . Exonuclease VIII of Escherichia coli I. Purification and physical properties . J. Biol. Chem. 258 : 10411 - 10417 . (10.1016/S0021-9258(17)44472-3) / J. Biol. Chem. / Exonuclease VIII of Escherichia coli I. Purification and physical properties by Joseph J. W. (1983)
  24. Kmiec , E. , and W. K. Holloman . 1981 . d protein of bacteriophage X promotes renaturation of DNA . J. Biol. Chem. 256 : 12636 - 12639 . (10.1016/S0021-9258(18)42938-9) / J. Biol. Chem. / d protein of bacteriophage X promotes renaturation of DNA by Kmiec E. (1981)
  25. Kolodner R. (Sidney Farber Cancer Institute Boston). 1992. Personal communication.
  26. Kushner , S. R. , H. Nagaishi , and A. J. Clarlk 1972 . Indirect suppression of recB and recC mutations by exonuclease I deficiency . Proc. Natl. Acad. Sci. USA 69 : 1366 - 1370 . (10.1073/pnas.69.6.1366) / Proc. Natl. Acad. Sci. USA / Indirect suppression of recB and recC mutations by exonuclease I deficiency by Kushner S. R. (1972)
  27. Kushner , S. R. , H. Nagaishi , and A. J. Clark . 1974 . Isolation of exonuclease VIII: the enzyme associated with the sbcA indirect suppressor . Proc. Natl. Acad. Sci. USA 71 : 3593 - 3597 . (10.1073/pnas.71.9.3593) / Proc. Natl. Acad. Sci. USA / Isolation of exonuclease VIII: the enzyme associated with the sbcA indirect suppressor by Kushner S. R. (1974)
  28. Luisi-DeLuca , C. , A. J. Clark , and R. D. Kolodner . 1988 . Analysis of the recE locus of Escherichia coli K-12 by use of polyclonal antibodies to exonuclease VIII . J. Bacteriol. 170 : 5797 - 5805 . (10.1128/jb.170.12.5797-5805.1988) / J. Bacteriol. / Analysis of the recE locus of Escherichia coli K-12 by use of polyclonal antibodies to exonuclease VIII by Luisi-DeLuca C. (1988)
  29. Luisi-DeLuca , C. , S. T. Lovett , and R. D. Kolodner . 1989 . Genetic and physical analysis of plasmid recombination in recB recC sbcB and recB recC sbcA Escherichia coli K-12 mutants . Genetics 122 : 269 - 278 . (10.1093/genetics/122.2.269) / Genetics / Genetic and physical analysis of plasmid recombination in recB recC sbcB and recB recC sbcA Escherichia coli K-12 mutants by Luisi-DeLuca C. (1989)
  30. Mahajan , S. K. , C. C. Chu , D. K. Willis , A. Templin , and A. J. Clark 1990 . Physical analysis of spontaneous and mutagen induced mutants of Escherichia coli K-12 expressing DNA exonuclease VIII activity . Genetics 125 : 261 - 273 . (10.1093/genetics/125.2.261) / Genetics / Physical analysis of spontaneous and mutagen induced mutants of Escherichia coli K-12 expressing DNA exonuclease VIII activity by Mahajan S. K. (1990)
  31. Mullis , K. B. , and F. A. Faloona . 1987 . Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction . Methods Enzymol. 155 : 335 - 350 . (10.1016/0076-6879(87)55023-6) / Methods Enzymol. / Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction by Mullis K. B. (1987)
  32. Nussbaum , A. , and A. Cohen . 1988 . Use of a bioluminescence gene reporter for the investigation of red-dependent and gam-dependent plasmid recombination in Escherichia coli K12 . J. Mol. Biol. 203 : 391 - 402 . (10.1016/0022-2836(88)90007-1) / J. Mol. Biol. / Use of a bioluminescence gene reporter for the investigation of red-dependent and gam-dependent plasmid recombination in Escherichia coli K12 by Nussbaum A. (1988)
  33. Radding , C. M. 1970 . The role of exonuclease and i protein of bacteriophage A in genetic recombination. I. Effects of red mutants on protein structure . J. Mol. Biol. 52 : 491 - 499 . (10.1016/0022-2836(70)90415-8) / J. Mol. Biol. / The role of exonuclease and i protein of bacteriophage A in genetic recombination. I. Effects of red mutants on protein structure by Radding C. M. (1970)
  34. Sandler , S. J. , and A. J. Clark 1990 . Factors affecting expression of the recF gene of E. coli K-12 . Gene 86 : 35 - 43 . (10.1016/0378-1119(90)90111-4) / Gene / Factors affecting expression of the recF gene of E. coli K-12 by Sandler S. J. (1990)
  35. Sanger , F. , S. Nicklen , and A. R. Coulson . 1977 . DNA sequencing with chain-terminating inhibitors . Proc. Natl. Acad. Sci. USA 74 : 5463 - 5467 . (10.1073/pnas.74.12.5463) / Proc. Natl. Acad. Sci. USA / DNA sequencing with chain-terminating inhibitors by Sanger F. (1977)
  36. Satin L. and A. J. Clark. Unpublished data.
  37. Shulman , M. J. , L. M. Hallick , H. Echols , and E. R. Signer . 1970 . Properties of recombination-deficient mutants of bacteriophage A . J. Mol. Biol. 52 : 501 - 520 . (10.1016/0022-2836(70)90416-X) / J. Mol. Biol. / Properties of recombination-deficient mutants of bacteriophage A by Shulman M. J. (1970)
  38. Sutcliffe , J. G. 1979 . Complete nucleotide sequence of the Escherichia coli plasmid pBR322 . Cold Spring Harbor Symp. Quant. Biol. 43 : 77 - 90 . (10.1101/SQB.1979.043.01.013) / Cold Spring Harbor Symp. Quant. Biol. / Complete nucleotide sequence of the Escherichia coli plasmid pBR322 by Sutcliffe J. G. (1979)
  39. Willis , D. K. , K. E. Fouts , S. D. Barbour , and A. J. Clark . 1983 . Physical characterization of transposon-induced mutations of the Rac prophage which affect expression and function of recE in Escherichia coli K-12 . J. Bacteriol. 156 : 727 - 736 . (10.1128/jb.156.2.727-736.1983) / J. Bacteriol. / Physical characterization of transposon-induced mutations of the Rac prophage which affect expression and function of recE in Escherichia coli K-12 by Willis D. K. (1983)
  40. Willis , D. K. , L. H. Satin , and A. J. Clark . 1985 . Mutationdependent suppression of recB21 recC22 by a region cloned from the Rac prophage of Escherichia coli K-12 . J. Bacteriol. 162 : 1166 - 1172 . (10.1128/jb.162.3.1166-1172.1985) / J. Bacteriol. / Mutationdependent suppression of recB21 recC22 by a region cloned from the Rac prophage of Escherichia coli K-12 by Willis D. K. (1985)
Dates
Type When
Created 8 years, 9 months ago (Nov. 7, 2016, 2:23 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:17 p.m.)
Indexed 1 year, 7 months ago (Jan. 18, 2024, 12:24 a.m.)
Issued 31 years, 9 months ago (Dec. 1, 1993)
Published 31 years, 9 months ago (Dec. 1, 1993)
Published Print 31 years, 9 months ago (Dec. 1, 1993)
Funders 0

None

@article{Clark_1993, title={Genetic and molecular analyses of the C-terminal region of the recE gene from the Rac prophage of Escherichia coli K-12 reveal the recT gene}, volume={175}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.175.23.7673-7682.1993}, DOI={10.1128/jb.175.23.7673-7682.1993}, number={23}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Clark, A J and Sharma, V and Brenowitz, S and Chu, C C and Sandler, S and Satin, L and Templin, A and Berger, I and Cohen, A}, year={1993}, month=dec, pages={7673–7682} }