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Nucleic Acids Research, 1990, Vol. 18, No. 15 4377-4384
© 1990

MOLECULAR BIOLOGY

Specificity of binding to four-way junctions in DNA by bacteriophage T7 endonuclease I

Carol A. Parsons and Stephen C. West*

Imperial Cancer Research Fund, Clare Hall Laboratories South Mimms, Herts EN6 3LD, UK

*To whom correspondence should be addressed

Received May 23, 1990. Revised July 5, 1990. Accepted July 5, 1990.

T7 endonuclease I binds specifically to four-way junctions in duplex DNA and promotes their resolution into linear duplexes. Under conditions in which the nuclease activity is blocked by the absence of divalent cations, the enzyme forms a distinct protein-DNA complex with the junction, as detected by gel retardation and filter binding assays. The formation of this complex is structure-specific and contrasts with the short-lived binding complexes formed on linear duplex DNA. The binding complex between T7 endonuclease I and a synthetic Holliday junction analog has been probed with hydroxyl radicals. The results indicate that the nuclease binds all four strands about the junction point.


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