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. 1992 Jun 25;20(12):3121–3125. doi: 10.1093/nar/20.12.3121

Joints formed by RecA protein from oligonucleotides and duplex DNA block initiation and elongation of transcription.

E I Golub 1, D C Ward 1, C M Radding 1
PMCID: PMC312447  PMID: 1620609

Abstract

In the presence of the non-hydrolyzable analog of ATP, ATP gamma S, RecA protein can polymerize on an oligodeoxy-ribonucleotide to form a stable oligonucleoprotein filament that can find its homologous sequence in double-stranded DNA. The homologous joint formed by the oligonucleotide and duplex DNA is stable only if RecA protein is not removed. Such a nucleoprotein joint, covering a part or all of the promoter region of T3 or T7 phage RNA polymerase, blocked transcription directed by those polymerases. The same kind of joint, located downstream of the RNA polymerase promoter, also inhibited elongation of transcription and caused accumulation of truncated transcripts. These observations suggest that RecA protein can be used to shut off transcription from any promoter of known sequence.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnott S., Selsing E. Letter: The structure of polydeoxyguanylic acid with polydeoxycytidylic acid. J Mol Biol. 1974 Sep 15;88(2):551–552. doi: 10.1016/0022-2836(74)90502-6. [DOI] [PubMed] [Google Scholar]
  2. Cheng S., Van Houten B., Gamper H. B., Sancar A., Hearst J. E. Use of psoralen-modified oligonucleotides to trap three-stranded RecA-DNA complexes and repair of these cross-linked complexes by ABC excinuclease. J Biol Chem. 1988 Oct 15;263(29):15110–15117. [PubMed] [Google Scholar]
  3. Cooney M., Czernuszewicz G., Postel E. H., Flint S. J., Hogan M. E. Site-specific oligonucleotide binding represses transcription of the human c-myc gene in vitro. Science. 1988 Jul 22;241(4864):456–459. doi: 10.1126/science.3293213. [DOI] [PubMed] [Google Scholar]
  4. Duval-Valentin G., Thuong N. T., Hélène C. Specific inhibition of transcription by triple helix-forming oligonucleotides. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):504–508. doi: 10.1073/pnas.89.2.504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ferrin L. J., Camerini-Otero R. D. Selective cleavage of human DNA: RecA-assisted restriction endonuclease (RARE) cleavage. Science. 1991 Dec 6;254(5037):1494–1497. doi: 10.1126/science.1962209. [DOI] [PubMed] [Google Scholar]
  6. Hanvey J. C., Shimizu M., Wells R. D. Intramolecular DNA triplexes in supercoiled plasmids. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6292–6296. doi: 10.1073/pnas.85.17.6292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Honigberg S. M., Gonda D. K., Flory J., Radding C. M. The pairing activity of stable nucleoprotein filaments made from recA protein, single-stranded DNA, and adenosine 5'-(gamma-thio)triphosphate. J Biol Chem. 1985 Sep 25;260(21):11845–11851. [PubMed] [Google Scholar]
  8. Howard-Flanders P., West S. C., Stasiak A. Role of RecA protein spiral filaments in genetic recombination. Nature. 1984 May 17;309(5965):215–219. doi: 10.1038/309215a0. [DOI] [PubMed] [Google Scholar]
  9. Hsieh P., Camerini-Otero C. S., Camerini-Otero R. D. Pairing of homologous DNA sequences by proteins: evidence for three-stranded DNA. Genes Dev. 1990 Nov;4(11):1951–1963. doi: 10.1101/gad.4.11.1951. [DOI] [PubMed] [Google Scholar]
  10. Htun H., Dahlberg J. E. Topology and formation of triple-stranded H-DNA. Science. 1989 Mar 24;243(4898):1571–1576. doi: 10.1126/science.2648571. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Postel E. H., Flint S. J., Kessler D. J., Hogan M. E. Evidence that a triplex-forming oligodeoxyribonucleotide binds to the c-myc promoter in HeLa cells, thereby reducing c-myc mRNA levels. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8227–8231. doi: 10.1073/pnas.88.18.8227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Radding C. M., Beattie K. L., Holloman W. K., Wiegand R. C. Uptake of homologous single-stranded fragments by superhelical DNA. IV. Branch migration. J Mol Biol. 1977 Nov;116(4):825–839. doi: 10.1016/0022-2836(77)90273-x. [DOI] [PubMed] [Google Scholar]
  14. Rajagopal P., Feigon J. Triple-strand formation in the homopurine:homopyrimidine DNA oligonucleotides d(G-A)4 and d(T-C)4. Nature. 1989 Jun 22;339(6226):637–640. doi: 10.1038/339637a0. [DOI] [PubMed] [Google Scholar]
  15. Rao B. J., Dutreix M., Radding C. M. Stable three-stranded DNA made by RecA protein. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):2984–2988. doi: 10.1073/pnas.88.8.2984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rao B. J., Jwang B., Radding C. M. RecA protein reinitiates strand exchange on isolated protein-free DNA intermediates. An ADP-resistant process. J Mol Biol. 1990 Jun 20;213(4):789–809. doi: 10.1016/S0022-2836(05)80264-5. [DOI] [PubMed] [Google Scholar]
  17. Rigas B., Welcher A. A., Ward D. C., Weissman S. M. Rapid plasmid library screening using RecA-coated biotinylated probes. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9591–9595. doi: 10.1073/pnas.83.24.9591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Young S. L., Krawczyk S. H., Matteucci M. D., Toole J. J. Triple helix formation inhibits transcription elongation in vitro. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10023–10026. doi: 10.1073/pnas.88.22.10023. [DOI] [PMC free article] [PubMed] [Google Scholar]

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