• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Sep 1991; 173(18): 5808–5821.
PMCID: PMC208314

Lambda Gam protein inhibits the helicase and chi-stimulated recombination activities of Escherichia coli RecBCD enzyme.

Abstract

The lambda Gam protein was isolated from cells containing a Gam-producing plasmid. The purified Gam protein was found to bind to RecBCD without displacing any of its subunits. Gam was shown to inhibit all known enzymatic activities of RecBCD: ATP-dependent single- and double-stranded DNA exonucleases, ATP-independent single-stranded endonuclease, and the ATP-dependent helicase. When produced in vivo, Gam inhibited chi-activated recombination in lambda red gam crosses but had little effect on the host's ability to act as a recipient in conjugational recombination. These experiments suggest that RecBCD possesses an additional "unknown" activity that is resistant to or induced by Gam. Additionally, the expression of Gam in recD mutants sensitizes the host to UV irradiation, indicating that Gam alters one or more of the in vivo activities of RecBC(D-).

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (3.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Amundsen SK, Neiman AM, Thibodeaux SM, Smith GR. Genetic dissection of the biochemical activities of RecBCD enzyme. Genetics. 1990 Sep;126(1):25–40. [PMC free article] [PubMed]
  • Amundsen SK, Taylor AF, Chaudhury AM, Smith GR. recD: the gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5558–5562. [PMC free article] [PubMed]
  • Barbour SD, Clark AJ. Biochemical and genetic studies of recombination proficiency in Escherichia coli. I. Enzymatic activity associated with recB+ and recC+ genes. Proc Natl Acad Sci U S A. 1970 Apr;65(4):955–961. [PMC free article] [PubMed]
  • Biek DP, Cohen SN. Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J Bacteriol. 1986 Aug;167(2):594–603. [PMC free article] [PubMed]
  • Bochner BR, Huang HC, Schieven GL, Ames BN. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brent R, Ptashne M. Mechanism of action of the lexA gene product. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4204–4208. [PMC free article] [PubMed]
  • Chalker AF, Leach DR, Lloyd RG. Escherichia coli sbcC mutants permit stable propagation of DNA replicons containing a long palindrome. Gene. 1988 Nov 15;71(1):201–205. [PubMed]
  • Chaudhury AM, Smith GR. Escherichia coli recBC deletion mutants. J Bacteriol. 1984 Nov;160(2):788–791. [PMC free article] [PubMed]
  • Chaudhury AM, Smith GR. A new class of Escherichia coli recBC mutants: implications for the role of RecBC enzyme in homologous recombination. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7850–7854. [PMC free article] [PubMed]
  • Chaudhury AM, Smith GR. Role of Escherichia coli RecBC enzyme in SOS induction. Mol Gen Genet. 1985;201(3):525–528. [PubMed]
  • Cohen A, Clark AJ. Synthesis of linear plasmid multimers in Escherichia coli K-12. J Bacteriol. 1986 Jul;167(1):327–335. [PMC free article] [PubMed]
  • Cox MM, Lehman IR. Enzymes of general recombination. Annu Rev Biochem. 1987;56:229–262. [PubMed]
  • Dykstra CC, Prasher D, Kushner SR. Physical and biochemical analysis of the cloned recB and recC genes of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):21–27. [PMC free article] [PubMed]
  • Finch PW, Storey A, Brown K, Hickson ID, Emmerson PT. Complete nucleotide sequence of recD, the structural gene for the alpha subunit of Exonuclease V of Escherichia coli. Nucleic Acids Res. 1986 Nov 11;14(21):8583–8594. [PMC free article] [PubMed]
  • Finch PW, Storey A, Chapman KE, Brown K, Hickson ID, Emmerson PT. Complete nucleotide sequence of the Escherichia coli recB gene. Nucleic Acids Res. 1986 Nov 11;14(21):8573–8582. [PMC free article] [PubMed]
  • Finch PW, Wilson RE, Brown K, Hickson ID, Tomkinson AE, Emmerson PT. Complete nucleotide sequence of the Escherichia coli recC gene and of the thyA-recC intergenic region. Nucleic Acids Res. 1986 Jun 11;14(11):4437–4451. [PMC free article] [PubMed]
  • Friedman SA, Hays JB. Selective inhibition of Escherichia coli recBC activities by plasmid-encoded GamS function of phage lambda. Gene. 1986;43(3):255–263. [PubMed]
  • Ganesan AK, Seawell PC. The effect of lexA and recF mutations on post-replication repair and DNA synthesis in Escherichia coli K-12. Mol Gen Genet. 1975 Dec 1;141(3):189–205. [PubMed]
  • Goldmark PJ, Linn S. An endonuclease activity from Escherichia coli absent from certain rec- strains. Proc Natl Acad Sci U S A. 1970 Sep;67(1):434–441. [PMC free article] [PubMed]
  • Hays JB, Smith TA, Friedman SA, Lee E, Coffman GL. RecF and RecBC function during recombination of nonreplicating, UV-irradiated phage lambda DNA and during other recombination processes. Cold Spring Harb Symp Quant Biol. 1984;49:475–483. [PubMed]
  • Horii Z, Clark AJ. Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K12: isolation and characterization of mutants. J Mol Biol. 1973 Oct 25;80(2):327–344. [PubMed]
  • Karu AE, Belk ED. Induction of E. coli recA protein via recBC and alternate pathways: quantitation by enzyme-linked immunosorbent assay (ELISA). Mol Gen Genet. 1982;185(2):275–282. [PubMed]
  • Karu AE, Sakaki Y, Echols H, Linn S. The gamma protein specified by bacteriophage gamma. Structure and inhibitory activity for the recBC enzyme of Escherichia coli. J Biol Chem. 1975 Sep 25;250(18):7377–7387. [PubMed]
  • Kulkarni SK, Stahl FW. Interaction between the sbcC gene of Escherichia coli and the gam gene of phage lambda. Genetics. 1989 Oct;123(2):249–253. [PMC free article] [PubMed]
  • Kushner SR. In vivo studies of temperature-sensitive recB and recC mutants. J Bacteriol. 1974 Dec;120(3):1213–1218. [PMC free article] [PubMed]
  • Kushner SR. Differential thermolability of exonuclease and endonuclease activities of the recBC nuclease isolated from thermosensitive recB and recC mutants. J Bacteriol. 1974 Dec;120(3):1219–1222. [PMC free article] [PubMed]
  • Lloyd RG, Buckman C. Identification and genetic analysis of sbcC mutations in commonly used recBC sbcB strains of Escherichia coli K-12. J Bacteriol. 1985 Nov;164(2):836–844. [PMC free article] [PubMed]
  • Lovett ST, Kolodner RD. Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2627–2631. [PMC free article] [PubMed]
  • Lovett ST, Luisi-DeLuca C, Kolodner RD. The genetic dependence of recombination in recD mutants of Escherichia coli. Genetics. 1988 Sep;120(1):37–45. [PMC free article] [PubMed]
  • Mackay V, Linn S. Selective inhibition of the dnase activity of the recBC enzyme by the DNA binding protein from Escherichia coli. J Biol Chem. 1976 Jun 25;251(12):3716–3719. [PubMed]
  • Mahan MJ, Roth JR. Role of recBC function in formation of chromosomal rearrangements: a two-step model for recombination. Genetics. 1989 Mar;121(3):433–443. [PMC free article] [PubMed]
  • McPartland A, Green L, Echols H. Control of recA gene RNA in E. coli: regulatory and signal genes. Cell. 1980 Jul;20(3):731–737. [PubMed]
  • Murphy KC, Casey L, Yannoutsos N, Poteete AR, Hendrix RW. Localization of a DNA-binding determinant in the bacteriophage P22 Erf protein. J Mol Biol. 1987 Mar 5;194(1):105–117. [PubMed]
  • Murphy KC, Fenton AC, Poteete AR. Sequence of the bacteriophage P22 anti-recBCD (abc) genes and properties of P22 abc region deletion mutants. Virology. 1987 Oct;160(2):456–464. [PubMed]
  • Naom IS, Morton SJ, Leach DR, Lloyd RG. Molecular organization of sbcC, a gene that affects genetic recombination and the viability of DNA palindromes in Escherichia coli K-12. Nucleic Acids Res. 1989 Oct 25;17(20):8033–8045. [PMC free article] [PubMed]
  • Niki H, Ogura T, Hiraga S. Linear multimer formation of plasmid DNA in Escherichia coli hopE (recD) mutants. Mol Gen Genet. 1990 Oct;224(1):1–9. [PubMed]
  • Palas KM, Kushner SR. Biochemical and physical characterization of exonuclease V from Escherichia coli. Comparison of the catalytic activities of the RecBC and RecBCD enzymes. J Biol Chem. 1990 Feb 25;265(6):3447–3454. [PubMed]
  • Ponticelli AS, Schultz DW, Taylor AF, Smith GR. Chi-dependent DNA strand cleavage by RecBC enzyme. Cell. 1985 May;41(1):145–151. [PubMed]
  • Poteete AR, Fenton AC. Lambda red-dependent growth and recombination of phage P22. Virology. 1984 Apr 15;134(1):161–167. [PubMed]
  • Poteete AR, Fenton AC, Murphy KC. Modulation of Escherichia coli RecBCD activity by the bacteriophage lambda Gam and P22 Abc functions. J Bacteriol. 1988 May;170(5):2012–2021. [PMC free article] [PubMed]
  • Poteete AR, Volkert MR. Activation of recF-dependent recombination in Escherichia coli by bacteriophage lambda- and P22-encoded functions. J Bacteriol. 1988 Sep;170(9):4379–4381. [PMC free article] [PubMed]
  • Roman LJ, Kowalczykowski SC. Characterization of the helicase activity of the Escherichia coli RecBCD enzyme using a novel helicase assay. Biochemistry. 1989 Apr 4;28(7):2863–2873. [PubMed]
  • Rothman RH, Clark AJ. The dependence of postreplication repair on uvrB in a recF mutant of Escherichia coli K-12. Mol Gen Genet. 1977 Oct 24;155(3):279–286. [PubMed]
  • Schägger H, von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. [PubMed]
  • Scharff R, Hanson MA, Hendler RW. A cellular factor involved in the formation of a DNA-synthesizing complex from DNA polymerase I in Escherichia coli. Biochim Biophys Acta. 1983 Apr 15;739(3):265–275. [PubMed]
  • Schultz DW, Taylor AF, Smith GR. Escherichia coli RecBC pseudorevertants lacking chi recombinational hotspot activity. J Bacteriol. 1983 Aug;155(2):664–680. [PMC free article] [PubMed]
  • Semerjian AV, Malloy DC, Poteete AR. Genetic structure of the bacteriophage P22 PL operon. J Mol Biol. 1989 May 5;207(1):1–13. [PubMed]
  • Sharma RC, Smith KC. Role of DNA polymerase I in postreplication repair: a reexamination with Escherichia coli delta polA. J Bacteriol. 1987 Oct;169(10):4559–4564. [PMC free article] [PubMed]
  • Smith GR. Homologous recombination in procaryotes. Microbiol Rev. 1988 Mar;52(1):1–28. [PMC free article] [PubMed]
  • Smith GR. Conjugational recombination in E. coli: myths and mechanisms. Cell. 1991 Jan 11;64(1):19–27. [PubMed]
  • Stahl FW, Stahl MM. Recombination pathway specificity of Chi. Genetics. 1977 Aug;86(4):715–725. [PMC free article] [PubMed]
  • Stahl FW, Thomason LC, Siddiqi I, Stahl MM. Further tests of a recombination model in which chi removes the RecD subunit from the RecBCD enzyme of Escherichia coli. Genetics. 1990 Nov;126(3):519–533. [PMC free article] [PubMed]
  • Syväoja JE. ATP-stimulated polymerase activity involving DNA polymerase I and a recB-dependent factor in extracts of Escherichia coli cells. Acta Chem Scand B. 1987 May;41(5):332–335. [PubMed]
  • Taylor A, Smith GR. Unwinding and rewinding of DNA by the RecBC enzyme. Cell. 1980 Nov;22(2 Pt 2):447–457. [PubMed]
  • Taylor AF, Smith GR. Substrate specificity of the DNA unwinding activity of the RecBC enzyme of Escherichia coli. J Mol Biol. 1985 Sep 20;185(2):431–443. [PubMed]
  • Taylor AF, Smith GR. Action of RecBCD enzyme on cruciform DNA. J Mol Biol. 1990 Jan 5;211(1):117–134. [PubMed]
  • Thaler DS, Sampson E, Siddiqi I, Rosenberg SM, Thomason LC, Stahl FW, Stahl MM. Recombination of bacteriophage lambda in recD mutants of Escherichia coli. Genome. 1989;31(1):53–67. [PubMed]
  • Unger RC, Clark AJ. Interaction of the recombination pathways of bacteriophage lambda and its host Escherichia coli K12: effects on exonuclease V activity. J Mol Biol. 1972 Oct 14;70(3):539–548. [PubMed]
  • Wang TC, Smith KC. Mechanisms for recF-dependent and recB-dependent pathways of postreplication repair in UV-irradiated Escherichia coli uvrB. J Bacteriol. 1983 Dec;156(3):1093–1098. [PMC free article] [PubMed]
  • Wang TC, Smith KC. Mechanism of sbcB-suppression of the recBC-deficiency in postreplication repair in UV-irradiated Escherichia coli K-12. Mol Gen Genet. 1985;201(2):186–191. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...