Abstract
The products of the recB and recC genes are necessary for conjugal recombination and for repair of chromosomal double-chain breaks in Escherichia coli. The recD gene product combines with the RecB and RecC proteins to comprise RecBCD enzyme but is required for neither recombination nor repair. On the contrary, RecBCD enzyme is an exonuclease that inhibits recombination by destroying linear DNA. The RecD ejection model proposes that RecBCD enzyme enters a DNA duplex at a double-chain end and travels destructively until it encounters the recombination hot spot sequence chi. Chi then alters the RecBCD enzyme by weakening the affinity of the RecD subunit for the RecBC heterodimer. With the loss of the RecD subunit, the resulting protein, RecBC(D-), becomes deficient for exonuclease activity and proficient as a recombinagenic helicase. To test the model, genetic crosses between lambda phage were conducted in cells containing chi on a nonhomologous plasmid. Upon delivering a double-chain break to the plasmid, lambda recombined as if the cells had become recD mutants. The ability of chi to alter lambda recombination in trans was reversed by overproducing the RecD subunit. These results indicate that chi can influence a recombination act without directly participating in it.
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- Amundsen S. K., Taylor A. F., Chaudhury A. M., Smith G. R. recD: the gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5558–5562. doi: 10.1073/pnas.83.15.5558. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chaudhury A. M., Smith G. R. 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. doi: 10.1073/pnas.81.24.7850. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheng K. C., Smith G. R. Distribution of Chi-stimulated recombinational exchanges and heteroduplex endpoints in phage lambda. Genetics. 1989 Sep;123(1):5–17. doi: 10.1093/genetics/123.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark A. J. Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet. 1973;7:67–86. doi: 10.1146/annurev.ge.07.120173.000435. [DOI] [PubMed] [Google Scholar]
- Dabert P., Ehrlich S. D., Gruss A. Chi sequence protects against RecBCD degradation of DNA in vivo. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12073–12077. doi: 10.1073/pnas.89.24.12073. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixon D. A., Churchill J. J., Kowalczykowski S. C. Reversible inactivation of the Escherichia coli RecBCD enzyme by the recombination hotspot chi in vitro: evidence for functional inactivation or loss of the RecD subunit. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):2980–2984. doi: 10.1073/pnas.91.8.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixon D. A., Kowalczykowski S. C. Homologous pairing in vitro stimulated by the recombination hotspot, Chi. Cell. 1991 Jul 26;66(2):361–371. doi: 10.1016/0092-8674(91)90625-9. [DOI] [PubMed] [Google Scholar]
- Dixon D. A., Kowalczykowski S. C. The recombination hotspot chi is a regulatory sequence that acts by attenuating the nuclease activity of the E. coli RecBCD enzyme. Cell. 1993 Apr 9;73(1):87–96. doi: 10.1016/0092-8674(93)90162-j. [DOI] [PubMed] [Google Scholar]
- Faulds D., Dower N., Stahl M. M., Stahl F. W. Orientation-dependent recombination hotspot activity in bacteriophage lambda. J Mol Biol. 1979 Jul 15;131(4):681–695. doi: 10.1016/0022-2836(79)90197-9. [DOI] [PubMed] [Google Scholar]
- Kobayashi I., Murialdo H., Crasemann J. M., Stahl M. M., Stahl F. W. Orientation of cohesive end site cos determines the active orientation of chi sequence in stimulating recA . recBC-mediated recombination in phage lambda lytic infections. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5981–5985. doi: 10.1073/pnas.79.19.5981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kobayashi I., Stahl M. M., Fairfield F. R., Stahl F. W. Coupling with packaging explains apparent nonreciprocality of Chi-stimulated recombination of bacteriophage lambda by RecA and RecBC functions. Genetics. 1984 Dec;108(4):773–794. doi: 10.1093/genetics/108.4.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kobayashi I., Stahl M. M., Stahl F. W. The mechanism of the chi-cos interaction in RecA-RecBC-mediated recombination in phage lambda. Cold Spring Harb Symp Quant Biol. 1984;49:497–506. doi: 10.1101/sqb.1984.049.01.056. [DOI] [PubMed] [Google Scholar]
- Kuzminov A., Schabtach E., Stahl F. W. Chi sites in combination with RecA protein increase the survival of linear DNA in Escherichia coli by inactivating exoV activity of RecBCD nuclease. EMBO J. 1994 Jun 15;13(12):2764–2776. doi: 10.1002/j.1460-2075.1994.tb06570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lam S. T., Stahl M. M., McMilin K. D., Stahl F. W. Rec-mediated recombinational hot spot activity in bacteriophage lambda. II. A mutation which causes hot spot activity. Genetics. 1974 Jul;77(3):425–433. doi: 10.1093/genetics/77.3.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Myers R. S., Stahl F. W. Chi and the RecBC D enzyme of Escherichia coli. Annu Rev Genet. 1994;28:49–70. doi: 10.1146/annurev.ge.28.120194.000405. [DOI] [PubMed] [Google Scholar]
- Sawitzke J. A., Stahl F. W. Phage lambda has an analog of Escherichia coli recO, recR and recF genes. Genetics. 1992 Jan;130(1):7–16. doi: 10.1093/genetics/130.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith G. R., Kunes S. M., Schultz D. W., Taylor A., Triman K. L. Structure of chi hotspots of generalized recombination. Cell. 1981 May;24(2):429–436. doi: 10.1016/0092-8674(81)90333-0. [DOI] [PubMed] [Google Scholar]
- Stahl F. W., McMilin K. D., Stahl M. M., Crasemann J. M., Lam S. The distribution of crossovers along unreplicated lambda bacteriophage chromosomes. Genetics. 1974 Jul;77(3):395–408. doi: 10.1093/genetics/77.3.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl F. W., Stahl M. M., Malone R. E., Crasemann J. M. Directionality and nonreciprocality of Chi-stimulated recombination in phage lambda. Genetics. 1980 Feb;94(2):235–248. doi: 10.1093/genetics/94.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl F. W., Stahl M. M. Recombination pathway specificity of Chi. Genetics. 1977 Aug;86(4):715–725. doi: 10.1093/genetics/86.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl F. W., Thomason L. C., Siddiqi I., Stahl M. M. 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. doi: 10.1093/genetics/126.3.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl M. M., Kobayashi I., Stahl F. W., Huntington S. K. Activation of Chi, a recombinator, by the action of an endonuclease at a distant site. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2310–2313. doi: 10.1073/pnas.80.8.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thaler D. S., Sampson E., Siddiqi I., Rosenberg S. M., Thomason L. C., Stahl F. W., Stahl M. M. Recombination of bacteriophage lambda in recD mutants of Escherichia coli. Genome. 1989;31(1):53–67. doi: 10.1139/g89-013. [DOI] [PubMed] [Google Scholar]