Abstract
When E. coli single-stranded DNA binding protein (SSB) coats single-stranded DNA (ssDNA) in the presence of 1 mM MgCl2 it inhibits the subsequent binding of recA protein, whereas SSB binding to ssDNA in 12 mM MgCl2 promotes the binding of recA protein. These two conditions correspond respectively to those which produce 'smooth' and 'beaded' forms of ssDNA-SSB filaments. By gel filtration and immunoprecipitation we observed active nucleoprotein filaments of recA protein and SSB on ssDNA that contained on average 1 monomer of recA protein per 4 nucleotides and 1 monomer of SSB per 20-22 nucleotides. Filaments in such a mixture, when digested with micrococcal nuclease produced a regular repeating pattern, approximately every 70-80 nucleotides, that differed from the pattern observed when only recA protein was bound to the ssDNA. We conclude that the beaded ssDNA-SSB nucleoprotein filament readily binds recA protein and forms an intermediate that is active in the formation of joint molecules and can retain substantially all of the SSB that was originally bound.
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- Beattie K. L., Wiegand R. C., Radding C. M. Uptake of homologous single-stranded fragments by superhelical DNA. II. Characterization of the reaction. J Mol Biol. 1977 Nov;116(4):783–803. doi: 10.1016/0022-2836(77)90271-6. [DOI] [PubMed] [Google Scholar]
- Bryant F. R., Taylor A. R., Lehman I. R. Interaction of the recA protein of Escherichia coli with single-stranded DNA. J Biol Chem. 1985 Jan 25;260(2):1196–1202. [PubMed] [Google Scholar]
- Bujalowski W., Lohman T. M. Escherichia coli single-strand binding protein forms multiple, distinct complexes with single-stranded DNA. Biochemistry. 1986 Dec 2;25(24):7799–7802. doi: 10.1021/bi00372a003. [DOI] [PubMed] [Google Scholar]
- Chase J. W., Murphy J. B., Whittier R. F., Lorensen E., Sninsky J. J. Amplification of ssb-1 mutant single-stranded DNA-binding protein in Escherichia coli. J Mol Biol. 1983 Feb 25;164(2):193–211. doi: 10.1016/0022-2836(83)90075-x. [DOI] [PubMed] [Google Scholar]
- Chase J. W., Williams K. R. Single-stranded DNA binding proteins required for DNA replication. Annu Rev Biochem. 1986;55:103–136. doi: 10.1146/annurev.bi.55.070186.000535. [DOI] [PubMed] [Google Scholar]
- Chrysogelos S., Griffith J. Escherichia coli single-strand binding protein organizes single-stranded DNA in nucleosome-like units. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5803–5807. doi: 10.1073/pnas.79.19.5803. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clore G. M., Gronenborn A. M., Greipel J., Maass G. Conformation of the DNA undecamer 5'd(A-A-G-T-G-T-G-A-T-A-T) bound to the single-stranded DNA binding protein of Escherichia coli. A time-dependent transferred nuclear Overhauser enhancement study. J Mol Biol. 1986 Jan 5;187(1):119–124. doi: 10.1016/0022-2836(86)90411-0. [DOI] [PubMed] [Google Scholar]
- Cox M. M., Lehman I. R. Enzymes of general recombination. Annu Rev Biochem. 1987;56:229–262. doi: 10.1146/annurev.bi.56.070187.001305. [DOI] [PubMed] [Google Scholar]
- Cox M. M., Lehman I. R. recA protein-promoted DNA strand exchange. Stable complexes of recA protein and single-stranded DNA formed in the presence of ATP and single-stranded DNA binding protein. J Biol Chem. 1982 Jul 25;257(14):8523–8532. [PubMed] [Google Scholar]
- Cunningham R. P., DasGupta C., Shibata T., Radding C. M. Homologous pairing in genetic recombination: recA protein makes joint molecules of gapped circular DNA and closed circular DNA. Cell. 1980 May;20(1):223–235. doi: 10.1016/0092-8674(80)90250-0. [DOI] [PubMed] [Google Scholar]
- Griffith J. D., Harris L. D., Register J., 3rd Visualization of SSB-ssDNA complexes active in the assembly of stable RecA-DNA filaments. Cold Spring Harb Symp Quant Biol. 1984;49:553–559. doi: 10.1101/sqb.1984.049.01.062. [DOI] [PubMed] [Google Scholar]
- Julin D. A., Riddles P. W., Lehman I. R. On the mechanism of pairing of single- and double-stranded DNA molecules by the recA and single-stranded DNA-binding proteins of Escherichia coli. J Biol Chem. 1986 Jan 25;261(3):1025–1030. [PubMed] [Google Scholar]
- Kahn R., Radding C. M. Separation of the presynaptic and synaptic phases of homologous pairing promoted by recA protein. J Biol Chem. 1984 Jun 25;259(12):7495–7503. [PubMed] [Google Scholar]
- Kowalczykowski S. C., Clow J., Somani R., Varghese A. Effects of the Escherichia coli SSB protein on the binding of Escherichia coli RecA protein to single-stranded DNA. Demonstration of competitive binding and the lack of a specific protein-protein interaction. J Mol Biol. 1987 Jan 5;193(1):81–95. doi: 10.1016/0022-2836(87)90629-2. [DOI] [PubMed] [Google Scholar]
- Leahy M. C., Radding C. M. Topography of the interaction of recA protein with single-stranded deoxyoligonucleotides. J Biol Chem. 1986 May 25;261(15):6954–6960. [PubMed] [Google Scholar]
- Lohman T. M., Bujalowski W., Overman L. B. E. coli single strand binding protein: a new look at helix-destabilizing proteins. Trends Biochem Sci. 1988 Jul;13(7):250–255. [PubMed] [Google Scholar]
- Lohman T. M., Overman L. B., Datta S. Salt-dependent changes in the DNA binding co-operativity of Escherichia coli single strand binding protein. J Mol Biol. 1986 Feb 20;187(4):603–615. doi: 10.1016/0022-2836(86)90338-4. [DOI] [PubMed] [Google Scholar]
- Lohman T. M., Overman L. B. Two binding modes in Escherichia coli single strand binding protein-single stranded DNA complexes. Modulation by NaCl concentration. J Biol Chem. 1985 Mar 25;260(6):3594–3603. [PubMed] [Google Scholar]
- McEntee K., Weinstock G. M., Lehman I. R. Binding of the recA protein of Escherichia coli to single- and double-stranded DNA. J Biol Chem. 1981 Aug 25;256(16):8835–8844. [PubMed] [Google Scholar]
- Menetski J. P., Kowalczykowski S. C. Interaction of recA protein with single-stranded DNA. Quantitative aspects of binding affinity modulation by nucleotide cofactors. J Mol Biol. 1985 Jan 20;181(2):281–295. doi: 10.1016/0022-2836(85)90092-0. [DOI] [PubMed] [Google Scholar]
- Morrical S. W., Lee J., Cox M. M. Continuous association of Escherichia coli single-stranded DNA binding protein with stable complexes of recA protein and single-stranded DNA. Biochemistry. 1986 Apr 8;25(7):1482–1494. doi: 10.1021/bi00355a003. [DOI] [PubMed] [Google Scholar]
- Muniyappa K., Shaner S. L., Tsang S. S., Radding C. M. Mechanism of the concerted action of recA protein and helix-destabilizing proteins in homologous recombination. Proc Natl Acad Sci U S A. 1984 May;81(9):2757–2761. doi: 10.1073/pnas.81.9.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neuendorf S. K., Cox M. M. Exchange of recA protein between adjacent recA protein-single-stranded DNA complexes. J Biol Chem. 1986 Jun 25;261(18):8276–8282. [PubMed] [Google Scholar]
- Overman L. B., Bujalowski W., Lohman T. M. Equilibrium binding of Escherichia coli single-strand binding protein to single-stranded nucleic acids in the (SSB)65 binding mode. Cation and anion effects and polynucleotide specificity. Biochemistry. 1988 Jan 12;27(1):456–471. doi: 10.1021/bi00401a067. [DOI] [PubMed] [Google Scholar]
- Prigodich R. V., Casas-Finet J., Williams K. R., Konigsberg W., Coleman J. E. 1H NMR (500 MHz) of gene 32 protein--oligonucleotide complexes. Biochemistry. 1984 Jan 31;23(3):522–529. doi: 10.1021/bi00298a019. [DOI] [PubMed] [Google Scholar]
- Radding C. M. Homologous pairing and strand exchange in genetic recombination. Annu Rev Genet. 1982;16:405–437. doi: 10.1146/annurev.ge.16.120182.002201. [DOI] [PubMed] [Google Scholar]
- Shibata T., Cunningham R. P., Radding C. M. Homologous pairing in genetic recombination. Purification and characterization of Escherichia coli recA protein. J Biol Chem. 1981 Jul 25;256(14):7557–7564. [PubMed] [Google Scholar]
- Tsang S. S., Muniyappa K., Azhderian E., Gonda D. K., Radding C. M., Flory J., Chase J. W. Intermediates in homologous pairing promoted by recA protein. Isolation and characterization of active presynaptic complexes. J Mol Biol. 1985 Sep 20;185(2):295–309. doi: 10.1016/0022-2836(85)90405-x. [DOI] [PubMed] [Google Scholar]
- West S. C., Cassuto E., Mursalim J., Howard-Flanders P. Recognition of duplex DNA containing single-stranded regions by recA protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2569–2573. doi: 10.1073/pnas.77.5.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]