PMID:17601790
| Citation |
Flanigan, A and Gardner, JF (2007) Interaction of the Gifsy-1 Xis protein with the Gifsy-1 attP sequence. J. Bacteriol. 189:6303-11 |
|---|---|
| Abstract |
The Gifsy-1 phage integrates site specifically into the Salmonella chromosome via an integrase-mediated site-specific recombination mechanism. Initial genetic analysis suggests that Gifsy-1 integrase-mediated excision of the Gifsy-1 phage is influenced by proteins encoded by both the Gifsy-1 and the Gifsy-2 phages. Our studies show that the Gifsy-1 Xis protein regulates the directionality of integrase-mediated excision of the Gifsy-1 phage. Electrophoretic mobility shift assays, DNase I footprinting, dimethyl sulfate (DMS) interference assays, and DMS protection assays were used to identify a 31-base-pair sequence in the attP region to which the Gifsy-1 protein binds. The results suggest that this recombination directionality factor binds in vitro to three imperfect direct repeats, spaced 10 base pairs apart, in a sequential and cooperative manner in the absence of other phage-encoded proteins. Our studies suggest that, while the Gifsy-1 Xis does not require additional factors for specific and high-affinity binding, it may form a microfilament on DNA similar to that described for the phage lambda Xis protein. |
| Links |
PubMed PMC1951908 Online version:10.1128/JB.00577-07 |
| Keywords |
Amino Acid Sequence; Attachment Sites, Microbiological; Base Sequence; DNA Footprinting; DNA, Bacterial/metabolism; DNA-Binding Proteins/metabolism; Electrophoretic Mobility Shift Assay; Molecular Sequence Data; Protein Binding; Salmonella Phages/genetics; Salmonella Phages/metabolism; Salmonella typhimurium/genetics; Salmonella typhimurium/virology; Sulfuric Acid Esters/metabolism; Viral Proteins/metabolism |
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