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Infection and Immunity, May 2000, p. 2457-2463, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The sigA Gene Which Is Borne on the she Pathogenicity Island of Shigella flexneri 2a Encodes an Exported Cytopathic Protease Involved in Intestinal Fluid Accumulation

Keith Al-Hasani,1 Ian R. Henderson,2 Harry Sakellaris,1 Kumar Rajakumar,1,3,* Travis Grant,3 James P. Nataro,2 Roy Robins-Browne,3 and Ben Adler1

Department of Microbiology, Monash University, Clayton, Victoria 3800,1 and Department of Microbiology and Infectious Diseases, Royal Children's Hospital, Parkville, Victoria 3052,3 Australia, and Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, Maryland 212012

Received 11 November 1999/Returned for modification 22 December 1999/Accepted 17 January 2000

In this study, the sigA gene situated on the she pathogenicity island of Shigella flexneri 2a was cloned and characterized. Sequence analysis showed that sigA encodes a 139.6-kDa protein which belongs to the SPATE (serine protease autotransporters of Enterobacteriaceae) subfamily of autotransporter proteins. The demonstration that SigA is autonomously secreted from the cell to yield a 103-kDa processed form and possesses a conserved C-terminal domain for export from the cell were consistent with the autotransporter pathway of secretion. Functional analysis showed that SigA is a secreted temperature-regulated serine protease capable of degrading casein. SigA was cytopathic for HEp-2 cells, suggesting that it may be a cell-altering toxin with a role in the pathogenesis of Shigella infections. SigA was at least partly responsible for the ability of S. flexneri to stimulate fluid accumulation in ligated rabbit ileal loops.

* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, Royal Children's Hospital, Parkville, Victoria 3052, Australia. Phone: 61-3-9344 2000. Fax: 61-3-934 55764. E-mail: Kumar.Rajakumar{at}med.monash.edu.au.

Infection and Immunity, May 2000, p. 2457-2463, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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