PMID: 9190821Jun 1, 1997Paper

Genetic analysis of second-site revertants of bacteriophage lambda integrase mutants

Journal of Bacteriology
Z WuJ F Gardner

Abstract

Bacteriophage lambda site-specific recombination is catalyzed by the phage-encoded integrase (Int) protein. Using a collection of 21 recombination-defective Int mutants, we performed a second-site reversion analysis. One of the primary mutants contained a valine-to-glutamic acid change at position 175 (V175E), and a pseudorevertant with a lysine change at this site (V175K) was also isolated. Relative to the wild-type protein, the V175E protein was defective in its ability to form the attL complex and to catalyze excision in vivo and in vitro. A mutant containing an alanine substitution (V175A) was made by site-directed mutagenesis, and it was more efficient than the V175K protein in forming the attL complex and promoting excision. These results indicate that a nonpolar side chain at residue 175 is required for function. The second primary mutant contained a proline-to-leucine change at position 243 (P243L). A true second-site revertant was isolated that contained a glutamic acid-to-lysine change (E218K). The P243L-E218K protein promoted recombination and bound arm-type sites more efficiently than the original P243L protein but not as efficiently as the protein containing the E218K substitution alone. The E218K substitution also...Continue Reading

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Citations

Jun 21, 2007·Proceedings of the National Academy of Sciences of the United States of America·Lauren M F MerloAntony M Dean
Aug 29, 2013·Biotechnology and Bioengineering·Thomas GajCarlos F Barbas
Oct 24, 2001·Journal of Bacteriology·N Messier, P H Roy
Dec 4, 2020·Nucleic Acids Research·Amer EliasMikhail Kolot

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