Abstract
Recombination between two different deletion alleles of a gene (neo) for neomycin and kanamycin resistance was studied in an Escherichia coli sbcA(-) recB(-)C(-) strain. The two homologous regions were in an inverted orientation on the same plasmid molecule. Kanamycin-resistant plasmids were selected and analyzed. The rate of recombination to form kanamycin-resistant plasmids was decreased by mutations in the recE, recF and recJ genes, but was not decreased by a mutation in the recA gene. It was found that these plasmids often possessed one wild-type kanamycin-resistant allele (neo(+)) while the other neo allele was still in its original (deletion) form. Among kanamycin-resistant plasmids with one wild-type and one parental allele it was often found that the region between the inverted repeats had been flipped (turned around) with respect to sites outside the inverted repeats. These results were interpreted as follows. Gene conversion, analogous to gene conversion in eukaryotic meiosis, is responsible for a unidirectional transfer of information from one neo deletion allele to the other. The flipping of the region between the inverted repeats is interpreted as analogous to the crossing over associated with gene conversion in eukaryotic meiosis. In contrast with a rec(+) strain, these products cannot be explained by two rounds of reciprocal crossing over involving a dimeric form as an intermediate. In the accompanying paper we present evidence that gene conversion by double-strand gap repair takes place in the same E. coli strain.
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- Amati P, Meselson M. Localized Negative Interference in Bacteriophage. Genetics. 1965 Mar;51(3):369–379. doi: 10.1093/genetics/51.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Boon T., Zinder N. D. A mechanism for genetic recombination generating one parent and one recombinant. Proc Natl Acad Sci U S A. 1969 Oct;64(2):573–577. doi: 10.1073/pnas.64.2.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doherty M. J., Morrison P. T., Kolodner R. Genetic recombination of bacterial plasmid DNA. Physical and genetic analysis of the products of plasmid recombination in Escherichia coli. J Mol Biol. 1983 Jul 5;167(3):539–560. doi: 10.1016/s0022-2836(83)80097-7. [DOI] [PubMed] [Google Scholar]
- Dohet C., Dzidić S., Wagner R., Radman M. Large non-homology in heteroduplex DNA is processed differently than single base pair mismatches. Mol Gen Genet. 1987 Jan;206(1):181–184. doi: 10.1007/BF00326556. [DOI] [PubMed] [Google Scholar]
- Gillen J. R., Willis D. K., Clark A. J. Genetic analysis of the RecE pathway of genetic recombination in Escherichia coli K-12. J Bacteriol. 1981 Jan;145(1):521–532. doi: 10.1128/jb.145.1.521-532.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kobayashi I., Ikeda H. Double Holliday structure: a possible in vivo intermediate form of general recombination in Escherichia coli. Mol Gen Genet. 1983;191(2):213–220. doi: 10.1007/BF00334816. [DOI] [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]
- Lieb M. Specific mismatch correction in bacteriophage lambda crosses by very short patch repair. Mol Gen Genet. 1983;191(1):118–125. doi: 10.1007/BF00330898. [DOI] [PubMed] [Google Scholar]
- Little J. W. An exonuclease induced by bacteriophage lambda. II. Nature of the enzymatic reaction. J Biol Chem. 1967 Feb 25;242(4):679–686. [PubMed] [Google Scholar]
- Lovett S. T., Clark A. J. Genetic analysis of the recJ gene of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):190–196. doi: 10.1128/jb.157.1.190-196.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
- Oishi M. An ATP-dependent deoxyribonuclease from Escherichia coli with a possible role in genetic recombination. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1292–1299. doi: 10.1073/pnas.64.4.1292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ponticelli A. S., Schultz D. W., Taylor A. F., Smith G. R. Chi-dependent DNA strand cleavage by RecBC enzyme. Cell. 1985 May;41(1):145–151. doi: 10.1016/0092-8674(85)90069-8. [DOI] [PubMed] [Google Scholar]
- Rossignol J. L., Nicolas A., Hamza H., Langin T. Origins of gene conversion and reciprocal exchange in Ascobolus. Cold Spring Harb Symp Quant Biol. 1984;49:13–21. doi: 10.1101/sqb.1984.049.01.004. [DOI] [PubMed] [Google Scholar]
- Sarthy P. V., Meselson M. Single burst study of rec- and red-mediated recombination in bacteriophage lambda. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4613–4617. doi: 10.1073/pnas.73.12.4613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shibata T., DasGupta C., Cunningham R. P., Radding C. M. Purified Escherichia coli recA protein catalyzes homologous pairing of superhelical DNA and single-stranded fragments. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1638–1642. doi: 10.1073/pnas.76.4.1638. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Silberstein Z., Cohen A. Synthesis of linear multimers of OriC and pBR322 derivatives in Escherichia coli K-12: role of recombination and replication functions. J Bacteriol. 1987 Jul;169(7):3131–3137. doi: 10.1128/jb.169.7.3131-3137.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
- Stahl F. W., Kobayashi I., Stahl M. M. Distance from cohesive end site cos determines the replication requirement for recombination in phage lambda. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6318–6321. doi: 10.1073/pnas.79.20.6318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl F. W., Kobayashi I., Stahl M. M. In phage lambda, cos is a recombinator in the red pathway. J Mol Biol. 1985 Jan 20;181(2):199–209. doi: 10.1016/0022-2836(85)90085-3. [DOI] [PubMed] [Google Scholar]
- Stahl F. W. Special sites in generalized recombination. Annu Rev Genet. 1979;13:7–24. doi: 10.1146/annurev.ge.13.120179.000255. [DOI] [PubMed] [Google Scholar]
- Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
- Thaler D. S., Stahl M. M., Stahl F. W. Double-chain-cut sites are recombination hotspots in the Red pathway of phage lambda. J Mol Biol. 1987 May 5;195(1):75–87. doi: 10.1016/0022-2836(87)90328-7. [DOI] [PubMed] [Google Scholar]
- Tomizawa J., Ogawa H. Breakage of DNA in rec+ and Rec- bacteria by disintegration of radiophosphorus atoms in DNA and possible cause of pleiotropic effects of RecA mutation. Cold Spring Harb Symp Quant Biol. 1968;33:243–251. doi: 10.1101/sqb.1968.033.01.028. [DOI] [PubMed] [Google Scholar]
- Tomizawa J., Ogawa H. Breakage of polynucleotide strands by disintegration of radiophosphorus atoms in DNA molecules and their repair. II. Simultaneous breakage of both strands. J Mol Biol. 1967 Nov 28;30(1):7–15. doi: 10.1016/0022-2836(67)90239-2. [DOI] [PubMed] [Google Scholar]
- Wildenberg J., Meselson M. Mismatch repair in heteroduplex DNA. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2202–2206. doi: 10.1073/pnas.72.6.2202. [DOI] [PMC free article] [PubMed] [Google Scholar]