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Nucleic Acids Research, 1988, Vol. 16, No. 21 10249-10266
© 1988

MOLECULAR BIOLOGY

Resolution of synthetic Holliday structures by an extract of human cells

Alan S. Waldman* and R. Michael Liskay

Departments of Therapeutic Radiology and Human Genetics, Yale University School of Medicine 333 Cedar Street, New Haven, CT 06510, USA

* To whom correspondence should be addressed

Received June 8, 1988. Accepted October 3, 1988.

Virtually all models for recombination between homologous DNA sequences invoke a branched intermediate known as a Holliday structure. The terminal steps of recombination are postulated to involve a specific cleavage through the four-way junction of a Holliday structure, in a process known as resolution. We have constructed a synthetic Holliday structure in which the position of the junction of the DNA duplexes can branch migrate through approximately 185 bp. Using this structure, we have found that a component of a cytoplasmic extract of Hela cells is capable of cleaving the central junction of the substrate in a manner consistent with resolution. The activity requires a divalent cation but does not require an exogenous energy source. This is the first reported resolution activity from a mammalian source.


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