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Journal of Bacteriology, November 2009, p. 6612-6617, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00628-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

A Novel Glucosyltransferase Involved in O-Antigen Modification of Shigella flexneri Serotype 1c{triangledown} ,{dagger}

Robert M. Stagg,1 Swee-Seong Tang,1,5 Nils I. A. Carlin,2 Kaisar A. Talukder,3 Phung D. Cam,4 and Naresh K. Verma1*

Biochemistry and Molecular Biology, Research School of Biology, The Australian National University, Canberra, ACT 0200, Australia,1 Department of Molecular Biology, SBL Vaccin AB, SE-105 21 Stockholm, Sweden,2 International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh,3 National Institute of Hygiene and Epidemiology, Department of Microbiology, Hanoi, Vietnam,4 University Malaya, Kuala Lumpur, Malaysia5

Received 13 May 2009/ Accepted 23 August 2009

The O antigen of serotype 1c differs from the unmodified O antigen of serotype Y by the addition of a disaccharide (two glucosyl groups) to the tetrasaccharide repeating unit. It was shown here that addition of the first glucosyl group is mediated by the previously characterized gtrI cluster, which is found within a cryptic prophage at the proA locus in the bacterial chromosome. Transposon mutagenesis was performed to disrupt the gene responsible for addition of the second glucosyl group, causing reversion to serotype 1a. Colony immunoblotting was used to identify the desired revertants, and subsequent sequencing, cloning, and functional expression successfully identified the gene encoding serotype 1c-specific O-antigen modification. This gene (designated gtrIC) was present as part of a three-gene cluster, similar to other S. flexneri glucosyltransferase genes. Relative to the other S. flexneri gtr clusters, the gtrIC cluster is more distantly related and appears to have arrived in S. flexneri from outside the species. Analysis of surrounding sequence suggests that the gtrIC cluster arrived via a novel bacteriophage that was subsequently rendered nonfunctional by a series of insertion events.

* Corresponding author. Present address: Biochemistry and Molecular Biology, Research School of Biology, The Australian National University, Canberra, ACT 0200, Australia. Phone: 61 2 6125 2666. Fax: 61 2 6125 0313. E-mail: naresh.verma{at}anu.edu.au

{triangledown} Published ahead of print on 28 August 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, November 2009, p. 6612-6617, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00628-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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