Abstract
The nucleoprotein filament formed by the RecA protein of Escherichia coli on single-stranded DNA catalyzes the hybridization of RNA transcripts with single-stranded DNA sequences at 37 degrees C, in vitro. RecA protein rapidly promotes hybridization, even when noncomplementary RNA is in a millionfold nucleotide excess over hybridizing RNA, and in a thousandfold nucleotide excess over hybridizing single-stranded DNA. Heterologous double-stranded DNA and RecA-coated noncomplementary single-stranded DNA are also poor competitors of RNA transcripts produced in vitro. Since large excesses of noncomplementary RNA fail to inhibit sharply the hybridization reaction by RecA protein under mild, non-degradative conditions, the reaction may be useful in the identification and isolation of transcripts produced in vivo.
MeSH Terms
DNA, Single-Stranded/metabolism
Electrophoresis
HeLa Cells
Humans
Nucleic Acid Hybridization/physiology
RNA, Messenger/metabolism
RNA, Ribosomal, 16S/metabolism
RNA, Ribosomal, 23S/metabolism
Rec A Recombinases/metabolism
Chemicals
DNA, Single-Stranded
RNA, Messenger
RNA, Ribosomal, 16S
RNA, Ribosomal, 23S
Rec A Recombinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kirkpatrick D P
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06510.
Radding C M
References (12)
12 references, click to expand
-
RNA-DNA hybridization promoted by E. coli RecA protein.
Nucleic Acids Res. 1992 Aug 25;20(16):4339-46
PMID: 1380698
-
Interaction of the recA protein of Escherichia coli with single-stranded DNA.
J Biol Chem. 1985 Jan 25;260(2):1196-202
PMID: 3838171
-
Kinetic complexity of RNA molecules.
J Mol Biol. 1972 Jan 14;63(1):21-39
PMID: 4622766
-
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
-
FORMATION AND PROPERTIES OF RNA-DNA COMPLEXES.
J Mol Biol. 1964 Jul;9:125-42
PMID: 14200377
-
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
-
A homogeneous nucleic acid hybridization assay based on strand displacement.
Nucleic Acids Res. 1987 Sep 11;15(17):6883-97
PMID: 3309890
-
Ability of RecA protein to promote a search for rare sequences in duplex DNA.
Proc Natl Acad Sci U S A. 1986 Dec;83(24):9586-90
PMID: 3467327
-
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
-
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
Nucleic Acids Res. 1984 Sep 25;12(18):7035-56
PMID: 6091052
-
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
-
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