Excision of a conjugative transposon in vitro by the Int and Xis proteins of Tn916.
The roles of purified Int and Xis proteins of the conjugative transposon Tn 916 in excision of a deletion derivative of the closely related element Tn 1545 were investigated. At a low salt concentration (37.5 mM NaCl), Int alone was able to promote limited excision to produce a covalently closed circular form of the transposon, showing that Tn 916 Int can catalyze both DNA cleavage and strand exchange. This reaction was stimulated by Xis. At higher salt concentrations (150 mM NaCl), excision by Int alone was reduced to barely detectable levels and Xis was required for excision. The low salt, Xis-stimulated reaction was approximately 8-fold more efficient than the high salt, Xis-dependent reaction. These results reflect in vivo requirements for Int and Xis in excision.
Rudy
C
C
Department of Microbiology and Immunology, Emory University School of Medicine, 1510 Clifton Road, Atlanta, GA 30322, USA.
Taylor
K L
KL
Hinerfeld
D
D
Scott
J R
JR
Churchward
G
G
D000595:Amino Acid Sequence; D003227:Conjugation, Genetic / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004251:DNA Transposable Elements; D013293:Enterococcus faecalis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D019426:Integrases / Q000235:genetics; D016133:Polymerase Chain Reaction; D012965:Sodium Chloride / Q000494:pharmacology; D013753:Tetracycline Resistance / Q000235:genetics*; D014764:Viral Proteins
D004251:DNA Transposable Elements; D014764:Viral Proteins; D012965:Sodium Chloride; D004247:DNA; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C063911:excisionase