Interaction of the Gifsy-1 Xis protein with the Gifsy-1 attP sequence.

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.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Flanigan A,Gardner JF

doi

10.1128/JB.00577-07

subject

Has Abstract

pub_date

2007-09-01 00:00:00

pages

6303-11

issue

17

eissn

0021-9193

issn

1098-5530

pii

JB.00577-07

journal_volume

189

pub_type

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