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Journal of Bacteriology, September 2007, p. 6303-6311, Vol. 189, No. 17
0021-9193/07/$08.00+0     doi:10.1128/JB.00577-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Interaction of the Gifsy-1 Xis Protein with the Gifsy-1 attP Sequence{triangledown}

Asa Flanigan1,2 and Jeffrey F. Gardner1*

University of Illinois at Urbana-Champaign, Department of Microbiology,1 College of Medicine, Urbana, Illinois2

Received 14 April 2007/ Accepted 18 June 2007

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.

* Corresponding author. Mailing address: Department of Microbiology, University of Illinois at Urbana-Champaign, 601 South Goodwin Avenue, Urbana, IL 61801. Phone: (217) 333-7287. Fax: (217) 244-6697. E-mail: jeffgard{at}life.uiuc.edu

{triangledown} Published ahead of print on 29 June 2007.

Journal of Bacteriology, September 2007, p. 6303-6311, Vol. 189, No. 17
0021-9193/07/$08.00+0     doi:10.1128/JB.00577-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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