Molecular and General Genetics MGG

, Volume 220, Issue 3, pp 456–460 | Cite as

RecA-ssDNA interaction: Induced strand cleavage by hydroxyl radical at a defined distance from the 5′ end

  • Eiko Akaboshi
  • Paul Howard-Flanders
Article
Received:

Summary

Interaction of the RecA protein with single-stranded DNA (ssDNA) was analyzed by challenge with the hydroxyl radical, which can cleave the DNA backbone. We found that RecA protein induces cleavage by the radical at a defined distance from the 5′ end. The cleavage was at the 11th nucleotide in many oligodeoxynucleotides. Cleavage may be intermittent since a second cleavage was induced at the 22nd or 21st site. This specific cleavage was observed under optimal conditions for filament formation, homologous pairing and strand exchange. Specificity in cleavage was, however, decreased by replacement of ATP by adenosine 5′-(γ-thio)triphosphate (ATPγS), replacement of RecA protein by a mutant (RecAl) protein, or an increase in Mg2– concentration. We propose that RecA protein induces a special structural alteration, such as bending, perhaps sequentially, on ssDNA and that this altered site plays an important role in homologous pairing and strand exchange.

Key words

DNA-protein interaction Iron(II) EDTA RecA 

Abbreviations

ssDNA

single-stranded DNA

dsDNA

doublestranded DNA

ATPγS

adenosine 5′-(γ-thio)triphosphate

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Eiko Akaboshi
    • 1
  • Paul Howard-Flanders
    • 1
  1. 1.Department of Molecular Biophysics and Biochemistry and Department of Therapeutic RadiologyYale UniversityNew HavenUSA

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