Abstract
The RecA protein of Escherichia coli catalyzes homologous pairing and strand exchange between a wide range of molecules showing nucleotide sequence complementarity, including a linear duplex and a single-stranded DNA molecule. We demonstrate that RecA can promote formation of joint molecules when the duplex contains an RNA/DNA hairpin and a single-stranded circle serves as the pairing partner. A chimeric RNA/DNA hairpin can be used to form stable joint molecules with as little as 15 bases of shared homology as long as the RNA stretch contains complementarity to the circle. The joint molecule bears some resemblance to a triple helical structure composed of RNA residues surrounded by two DNA strands which are in a parallel orientation. Evidence is presented that supports the notion that short stretches of RNA can be used in homologous pairing reactions at lengths below that required for DNA-DNA heteroduplex formation.
MeSH Terms
Bacteriophage M13
Base Composition
Base Sequence
DNA, Single-Stranded/chemistry,metabolism
DNA, Viral/chemistry,metabolism
Escherichia coli/genetics,metabolism
Molecular Sequence Data
Nucleic Acid Conformation
Oligodeoxyribonucleotides
RNA, Bacterial/chemistry,metabolism
Rec A Recombinases/metabolism
Ribonucleases
Sequence Homology, Nucleic Acid
Chemicals
DNA, Single-Stranded
DNA, Viral
Oligodeoxyribonucleotides
RNA, Bacterial
Rec A Recombinases
Ribonucleases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kotani H
Department of Pharmacology, Jefferson Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Germann M W
Andrus A
Vinayak R
Mullah B
Kmiec E B
References (20)
20 references, click to expand
-
Biochemical and biological function of Escherichia coli RecA protein: behavior of mutant RecA proteins.
Biochimie. 1991 Feb-Mar;73(2-3):289-304
PMID: 1883888
-
A role for RNA synthesis in homologous pairing events.
Mol Cell Biol. 1994 Sep;14(9):6097-106
PMID: 7520527
-
By searching processively RecA protein pairs DNA molecules that share a limited stretch of homology.
Cell. 1983 Sep;34(2):647-54
PMID: 6616624
-
Homologous recognition and triplex formation promoted by RecA protein between duplex oligonucleotides and single-stranded DNA.
J Mol Biol. 1993 Jan 20;229(2):328-43
PMID: 8381491
-
A nuclease from mung bean sprouts.
J Biol Chem. 1962 Feb;237:506-11
PMID: 13918439
-
Transcription activates RecA-promoted homologous pairing of nucleosomal DNA.
Mol Cell Biol. 1994 Mar;14(3):1949-55
PMID: 8114726
-
Conformational influence of the ribose 2'-hydroxyl group: crystal structures of DNA-RNA chimeric duplexes.
Biochemistry. 1993 Apr 6;32(13):3221-37
PMID: 7681688
-
Genetic recombination of nucleosomal templates is mediated by transcription.
Mol Gen Genet. 1994 Aug 15;244(4):410-9
PMID: 8078467
-
The synapsis event in the homologous pairing of DNAs: RecA recognizes and pairs less than one helical repeat of DNA.
Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6492-6
PMID: 1631148
-
Exonuclease VII of Escherichia coli. Mechanism of action.
J Biol Chem. 1974 Jul 25;249(14):4553-61
PMID: 4602030
-
The RecA protein: structure and function.
Crit Rev Biochem Mol Biol. 1990;25(6):415-56
PMID: 2292186
-
Pairing of homologous DNA sequences by proteins: evidence for three-stranded DNA.
Genes Dev. 1990 Nov;4(11):1951-63
PMID: 2276627
-
How specific is the first recognition step of homologous recombination?
Trends Biochem Sci. 1995 Mar;20(3):109-13
PMID: 7709428
-
Specificity of immunoglobulin heavy chain switch correlates with activity of germline heavy chain genes prior to switching.
EMBO J. 1986 Jan;5(1):95-102
PMID: 3007121
-
Stability and properties of double and triple helices: dramatic effects of RNA or DNA backbone composition.
Science. 1992 Nov 27;258(5087):1463-6
PMID: 1279808
-
Structure of a DNA:RNA hybrid duplex. Why RNase H does not cleave pure RNA.
J Mol Biol. 1993 Oct 5;233(3):509-23
PMID: 8411159
-
Structure and expression of germ line immunoglobulin gamma 2b transcripts.
Mol Cell Biol. 1988 Apr;8(4):1849-52
PMID: 3132612
-
Enzymes of general recombination.
Annu Rev Biochem. 1987;56:229-62
PMID: 3304134
-
Genetic rearrangements and gene amplification in Escherichia coli: DNA sequences at the junctures of amplified gene fusions.
Genes Dev. 1987 May;1(3):227-37
PMID: 3315849
-
Interactions of three strands in joints made by RecA protein.
Biochemistry. 1993 Dec 7;32(48):13146-55
PMID: 8241169