The synaptic complex of RecA protein participates in hybridization and inverse strand exchange reactions

Howard B. Gamper, Christopher J. Nulf, David R. Corey, Eric B. Kmiec

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

RecA protein catalyzes strand exchange between homologous single-stranded and doublestranded DNAs. In the presence of ATPγS, the post-strand exchange synaptic complex is a stable end product that can be studied. Here we ask whether such complexes can hybridize to or exchange with DNA, 2′-OMe RNA, PNA, or LNA oligonucleotides. Using a gel mobility shift assay, we show that the displaced strand of a 45 bp synaptic complex can hybridize to complementary oligonucleotides with different backbones to form a four-stranded (double D-loop) joint that survives removal of the RecA protein. This hybridization reaction, which confirms the single-stranded character of the displaced strand in a synaptic complex, might initiate recombination-dependent DNA replication if it occurs in vivo. We also show that either strand of the heteroduplex in a 30 bp synaptic complex can be replaced with a homologous DNA oligonucleotide in a strand exchange reaction that is mediated by the RecA filament. Consistent with the important role that deoxyribose plays in strand exchange, oligonucleotides with non-DNA backbones did not participate in this reaction. The hybridization and strand exchange reactions reported here demonstrate that short synaptic complexes are dynamic structures even in the presence of ATPγS.

Original languageEnglish (US)
Pages (from-to)2643-2655
Number of pages13
JournalBiochemistry
Volume42
Issue number9
DOIs
StatePublished - Mar 11 2003

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Rec A Recombinases
Oligonucleotides
DNA
Deoxyribose
Single-Stranded DNA
Electrophoretic Mobility Shift Assay
DNA Replication
Genetic Recombination
Assays
Joints
Gels
RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

The synaptic complex of RecA protein participates in hybridization and inverse strand exchange reactions. / Gamper, Howard B.; Nulf, Christopher J.; Corey, David R.; Kmiec, Eric B.

In: Biochemistry, Vol. 42, No. 9, 11.03.2003, p. 2643-2655.

Research output: Contribution to journalArticle

Gamper, Howard B. ; Nulf, Christopher J. ; Corey, David R. ; Kmiec, Eric B. / The synaptic complex of RecA protein participates in hybridization and inverse strand exchange reactions. In: Biochemistry. 2003 ; Vol. 42, No. 9. pp. 2643-2655.
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