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Nucleic Acids Research, 1990, Vol. 18, No. 13 3953-3959
© 1990

MOLECULAR BIOLOGY

A comparison of the effects of single-base and triple-base changes in the integrase arm-type binding sites on the site-specific recombination of bacteriophage lambda

Thomas E. Numrych, Richard I. Gumport1 and Jeffrey F. Gardner*

Department of Microbiology, University of Illinois Urbana, IL 61801, USA 1Department of Biochemistry and College of Medicine, University of Illinois Urbana, IL 61801, USA

*To whom correspondence should be addressed at Department of Microbiology, 131 Burrill Hall, 407 S. Goodwin, Urbana, IL 61801, USA

Triple-base changes were made in each of the five Integrase (Int) arm-type binding sites of bacteriophage lambda. These triple changes, called ten mutants, were compared with single-base changes (hen mutants) for their effects on integrative and excisive recombination. The presence of ten or hen mutations in the P1, P'2, or P'3 sites inhibited integration, but the ten P'3 mutant was 10-fold more defective than the analagous hen mutant. The results with these mutants suggest that the P1, P'2, P'3, and possibly the P'1 sites are required for integration. In wild-type E. coli, the ten P'1 mutant reduced the frequency of excision 5-fold, whereas the hen P'1 mutant had no effect. The presence of ten mutations in the P2, P'1, or P'2 sites inhibited {lambda} excision in an E. coli strain deficient in the production of FIS, while hen mutations in the P2 and P'2 sites had little or no effect. The results with the ten mutants suggest that the P2, P'1, and P'2 sites are required for excision. The differences in the severity of the effects between the ten and hen mutations may be due to the inability of cooperative interactions among Int, IHF, Xis, and FIS to overcome the disruption of Int binding to sites with triple-base changes compared to sites with single-base changes.


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