Complete DNA Sequence and Analysis of the Large Virulence Plasmid of Shigella flexneri

doi:10.1128/IAI.69.5.3271-3285.2001

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Infection and Immunity, May 2001, p. 3271-3285, Vol. 69, No. 5
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.5.3271-3285.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Complete DNA Sequence and Analysis of the Large Virulence Plasmid of Shigella flexneri

Malabi M. Venkatesan,¹^,^* Marcia B. Goldberg,²^,^* Debra J. Rose,³ Erik J. Grotbeck,³ Valerie Burland,³ and Frederick R. Blattner³

Department of Enteric Infections, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910¹; Infectious Disease Division, Massachusetts General Hospital, Boston, Massachuetts 02114²; and Laboratory of Genetics, University of Wisconsin, Madison, Wisconsin 53706³

Received 19 September 2000/Returned for modification 22 November 2000/Accepted 31 January 2001

The complete sequence analysis of the 210-kb Shigella flexneri 5a virulence plasmid was determined. Shigella spp. cause dysenteryand diarrhea by invasion and spread through the colonic mucosa.Most of the known Shigella virulence determinants are encodedon a large plasmid that is unique to virulent strains of Shigellaand enteroinvasive Escherichia coli; these known genes accountfor approximately 30 to 35% of the virulence plasmid. In the completesequence of the virulence plasmid, 286 open reading frames (ORFs)were identified. An astonishing 153 (53%) of these were relatedto known and putative insertion sequence (IS) elements; no knownbacterial plasmid has previously been described with such a highproportion of IS elements. Four new IS elements were identified.Fifty putative proteins show no significant homology to proteinsof known function; of these, 18 have a G+C content of less than40%, typical of known virulence genes on the plasmid. These 18constitute potentially unknown virulence genes. Two alleles ofshet2 and five alleles of ipaH were also identified on the plasmid.Thus, the plasmid sequence suggests a remarkable history of IS-mediatedacquisition of DNA across bacterial species. The complete sequencewill permit targeted characterization of potential new Shigellavirulencedeterminants.

^* Corresponding author. Mailing address for Malabi M. Venkatesan: Department of Enteric Infections, Division of CommunicableDiseases and Immunology, Walter Reed Army Institute of Research,503 Robert Forney Dr., Silver Spring, MD 20910. Phone: (301) 319-9764.Fax: (301) 319-9801. E-mail: Malabi.Venkatesan{at}NA.AMEDD.ARMY.MIL.Mailing address for Marcia B. Goldberg: Infectious Disease Division,Massachusetts General Hospital, GRJ504, 55 Fruit St., Boston,MA 02114. Phone: (617) 432-3238. Fax: (617) 432-3259. E-mail:mgoldberg1{at}partners.org.

Infection and Immunity, May 2001, p. 3271-3285, Vol. 69, No. 5
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.5.3271-3285.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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