Mutagenesis of the IS1 transposase: importance of a His-Arg-Tyr triad for activity.
Inspection of the primary sequence of the IS1 transposase suggested that it carries residues which are characteristic of the active site of integrases of the bacteriophage lambda family (Int). In particular, these include a highly conserved triad: His-Arg-Tyr. The properties of mutants made at each of these positions were investigated in vivo. The results of several different assays confirm that each is important for transposase activity. Moreover, as in the case of members of the Int family, different mutations of the His residue exhibited different effects. In a particular, His-to-Leu mutation resulted in complete inactivation whereas the equivalent His-to-Gln mutation retained low but significant levels of activity.
Serre
M C
MC
Laboratoire de Microbiologie et Génétique Moléculaire, Centre National de la Recherche Scientifique, Toulouse, France.
Turlan
C
C
Bortolin
M
M
Chandler
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009024:Morphogenesis; D016297:Mutagenesis, Site-Directed; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D013014:SOS Response, Genetics / Q000235:genetics; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D019895:Transposases
D009713:Nucleotidyltransferases; D019895:Transposases