Home LiteratureArticle Details
PMID: 1380698 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

RNA-DNA hybridization promoted by E. coli RecA protein.

Nucleic acids research ·Vol. 20 ·No. 16 ·1992-08-25 ·Pages 4339-46

Kirkpatrick DP, Rao BJ, Radding CM

Abstract

RecA protein of E. coli plays a central regulatory role that is induced by damage to DNA and results in the inactivation of LexA repressor. In vitro, RecA protein binds preferentially to single-stranded DNA to form a nucleoprotein filament that can recognize homology in naked duplex DNA and promote extensive strand exchange. Although RecA protein shows little tendency at neutral pH to bind to RNA, we found that it nonetheless catalyzed at 37 degrees C the hybridization of complementary RNA and single-stranded DNA sequences. Hybrids made by RecA protein at 37 degrees C appeared indistinguishable from ones prepared by thermal annealing. RNA-DNA hybridization by RecA protein at neutral pH required, as does RecA-promoted homologous pairing, optimal conditions for the formation of RecA nucleoprotein filaments. The cosedimentation of RNA with those filaments further paralleled observations made on the formation of networks of nucleoprotein filaments with double-stranded DNA, an instrumental intermediate in homologous pairing in vitro. These similarities with the pairing reaction support the view that RecA protein acts specifically in the hybridization reaction.

MeSH Terms
Bacterial Proteins/metabolism DNA, Single-Stranded/metabolism Electrophoresis Escherichia coli/genetics Hydrogen-Ion Concentration Magnesium/metabolism Nucleic Acid Hybridization/physiology RNA/metabolism Rec A Recombinases/metabolism Temperature
Chemicals
Bacterial Proteins DNA, Single-Stranded RNA Rec A Recombinases Magnesium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kirkpatrick D P
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06510.
Rao B J
Radding C M
References (33)
33 references, click to expand
  1. Activation of protease-constitutive recA proteins of Escherichia coli by all of the common nucleoside triphosphates.
    J Bacteriol. 1988 Oct;170(10):4816-22 PMID: 3049549
  2. The mechanism of the search for homology promoted by recA protein. Facilitated diffusion within nucleoprotein networks.
    J Biol Chem. 1986 Oct 5;261(28):13087-96 PMID: 3020024
  3. Capacity of RecA protein to bind preferentially to UV lesions and inhibit the editing subunit (epsilon) of DNA polymerase III: a possible mechanism for SOS-induced targeted mutagenesis.
    Proc Natl Acad Sci U S A. 1986 Feb;83(3):619-23 PMID: 3456159
  4. Linear dichroism study of RecA-DNA complexes. Structural evidence and binding stoichiometries.
    J Biol Chem. 1987 Jun 15;262(17):8109-11 PMID: 3597365
  5. 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 PMID: 4057248
  6. The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication.
    Gene. 1981 Sep;14(4):289-99 PMID: 6271628
  7. 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):2757-61 PMID: 6326142
  8. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  9. E. coli recA protein-directed cleavage of phage lambda repressor requires polynucleotide.
    Nature. 1980 Jan 3;283(5742):26-30 PMID: 6444245
  10. Separation of the presynaptic and synaptic phases of homologous pairing promoted by recA protein.
    J Biol Chem. 1984 Jun 25;259(12):7495-503 PMID: 6539775
  11. Isolation and visualization of active presynaptic filaments of recA protein and single-stranded DNA.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):7026-30 PMID: 6594678
  12. Binding of the recA protein of Escherichia coli to single- and double-stranded DNA.
    J Biol Chem. 1981 Aug 25;256(16):8835-44 PMID: 7021553
  13. Homologous pairing in genetic recombination: recA protein makes joint molecules of gapped circular DNA and closed circular DNA.
    Cell. 1980 May;20(1):223-35 PMID: 7388943
  14. Characteristics of purified recA protein and the regulation of its synthesis in vivo.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 2:909-15 PMID: 158476
  15. Nature of the SOS-inducing signal in Escherichia coli. The involvement of DNA replication.
    J Mol Biol. 1990 Mar 5;212(1):79-96 PMID: 2108251
  16. Active nucleoprotein filaments of single-stranded binding protein and recA protein on single-stranded DNA have a regular repeating structure.
    Nucleic Acids Res. 1990 Jul 11;18(13):3967-73 PMID: 2374716
  17. Binding stoichiometry and structure of RecA-DNA complexes studied by flow linear dichroism and fluorescence spectroscopy. Evidence for multiple heterogeneous DNA co-ordination.
    J Mol Biol. 1989 Jan 5;205(1):137-47 PMID: 2926802
  18. Kinetics of DNA renaturation catalyzed by the RecA protein of Escherichia coli.
    Biochemistry. 1985 Jul 30;24(16):4345-51 PMID: 2996595
  19. Networks of DNA and RecA protein are intermediates in homologous pairing.
    Biochemistry. 1985 Jun 18;24(13):3226-32 PMID: 3161539
  20. A homogeneous nucleic acid hybridization assay based on strand displacement.
    Nucleic Acids Res. 1987 Sep 11;15(17):6883-97 PMID: 3309890
  21. 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):11845-51 PMID: 3840165
  22. Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325.
    Gene. 1980 May;9(3-4):287-305 PMID: 6248430
  23. Homologous pairing and strand exchange in genetic recombination.
    Annu Rev Genet. 1982;16:405-37 PMID: 6297377
  24. Studies of the mechanism of DNA pairing by the RecA protein of Escherichia coli.
    Cold Spring Harb Symp Quant Biol. 1984;49:535-9 PMID: 6397308
  25. Homologous pairing in genetic recombination. Purification and characterization of Escherichia coli recA protein.
    J Biol Chem. 1981 Jul 25;256(14):7557-64 PMID: 6454691
  26. Homologous pairing in genetic recombination. The pairing reaction catalyzed by Escherichia coli recA protein.
    J Biol Chem. 1981 Jul 25;256(14):7565-72 PMID: 7019209
  27. The SOS regulatory system of Escherichia coli.
    Cell. 1982 May;29(1):11-22 PMID: 7049397
  28. RecA-mediated annealing of single-stranded DNA and its relation to the mechanism of homologous recombination.
    J Mol Biol. 1991 Sep 5;221(1):131-45 PMID: 1920401
  29. The RecA protein: structure and function.
    Crit Rev Biochem Mol Biol. 1990;25(6):415-56 PMID: 2292186
  30. Characterization of the DNA binding activity of stable RecA-DNA complexes. Interaction between the two DNA binding sites within RecA helical filaments.
    J Mol Biol. 1990 Mar 5;212(1):97-112 PMID: 2319601
  31. Activation of protease-constitutive recA proteins of Escherichia coli by rRNA and tRNA.
    J Bacteriol. 1988 Oct;170(10):4823-7 PMID: 2459110
  32. On the mechanism of renaturation of complementary DNA strands by the recA protein of Escherichia coli.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):297-301 PMID: 2982147
  33. Specific and cooperative binding of E. coli single-stranded DNA binding protein to mRNA.
    Nucleic Acids Res. 1987 Jul 10;15(13):5241-50 PMID: 3299265
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1992-08-25
Pages
4339-46
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334145
Subset
IM
Grants
NHGRI NIH HHS · HG00338-02 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]