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Nucleic Acids Research, 1989, Vol. 17, No. 5 2005-2022
© 1989

MOLECULAR BIOLOGY

Integration host factor (IHF) stimulates binding of the gpNu1 subunit of {lambda} terminase to cos DNA

Gayle Shinder and Marvin Gold

Department of Medical Genetics, University of Toronto Toronto, Ontario M5S 1A8, Canada

Received October 21, 1988. Revised January 31, 1989. Accepted January 31, 1989.

The {lambda} terminase enzyme binds to the cohesive end sites (cos) of multimeric replicating {lambda} DNA and introduces staggered nicks to regenerate the 12 bp single-stranded cohesive ends of the mature phage genome.In vitro this endonucleolytic cleavage requires spermidine, magnesium ions, ATP and a host factor. One of the E. coli proteins which can fulfill this latter requirement is integration Host Factor (IHF). IHF and the gpNul subunit of terminase can bind simultaneously to their own specific binding sites at cos. DNase I footprinting experiments suggest that IHF may promote gpNu1 binding. Although no specific gpNu1 binding to the left side of cos can be detected, this DNA segment does play a specific role since a cos fragment that does not include the left side or whose left side is replaced 1:y non-cos sequences, is unable to bind gpNu1 unless either spermidine or IHF is present. Binding studies on the right side of cos using individual or combinations of gpNu1 binding sites I, II and III indicate that binding at sites I and II is not optimal unless site III is present.


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