Gene 4 helicase of bacteriophage T7 mediates strand transfer through pyrimidine dimers, mismatches, and nonhomologous regions

  1. Daochun Kong*,
  2. Jack D. Griffith, and
  3. Charles C. Richardson*,
  1. *Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599

Abstract

In bacteriophage T7 the gene 2.5 single-stranded DNA-binding protein and the gene 4 helicase together promote the annealing of homologous regions of two DNA partners to form a joint molecule and subsequent strand transfer. In this reaction T7 gene 2.5 protein is essential for joint molecule formation, but is not required for T7 gene 4 protein-mediated strand transfer. T7 gene 4 helicase alone is able to mediate strand transfer, provided that a joint molecule is available. The present paper shows that, in addition, strand transfer proceeds at a normal rate even when both DNA partners contain ultraviolet-induced pyrimidine dimers (0.6 dimer per 100 nt). An insert of a relatively long (842-nt) segment of nonhomologous DNA in the single-stranded DNA partner has no effect on strand transfer, whereas its presence in the double-stranded partner prevents strand transfer. A short insert (37 nt) can be tolerated in either partner. Thus, DNA helicase is able to participate in recombinational DNA repair through its role in strand exchange, providing a pathway distinct from nucleotide excision repair.

Footnotes

  • To whom reprint requests should be addressed. e-mail: ccr{at}bcmp.med.harvard.edu.

  • Charles C. Richardson

  • ABBREVIATIONS:
    ssDNA,
    single-stranded DNA;
    dsDNA,
    double-stranded DNA;
    SSB protein,
    ssDNA-binding protein;
    RF1,
    replicative form 1
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