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Journal of Bacteriology, July 2003, p. 3764-3772, Vol. 185, No. 13
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.13.3764-3772.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Cluster of Type IV Secretion Genes in Helicobacter pylori's Plasticity Zone

Dangeruta Kersulyte,1 Billie Velapatiño,1,2 Asish K. Mukhopadhyay,1,{dagger} Lizbeth Cahuayme,2 Alejandro Bussalleu,3 Juan Combe,3 Robert H. Gilman,2 and Douglas E. Berg1*

Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110,1 Department of Microbiology,2 Department of Gastroenterology, Facultad de Medecina, Universidad Peruana Cayetano Heredia, Lima 31, Peru3

Received 21 February 2003/ Accepted 21 April 2003

Some genes present in only certain strains of the genetically diverse gastric pathogen Helicobacter pylori may affect its phenotype and/or evolutionary potential. Here we describe a new 16.3-kb segment, 7 of whose 16 open reading frames are homologs of type IV secretion genes (virB4, virB7 to virB11, and virD4), the third such putative secretion gene cluster found in H. pylori. This segment, to be called tfs3, was discovered by subtractive hybridization and chromosome walking. Full-length and truncated tfs3 elements were found in 20 and 19%, respectively, of 94 strains tested, which were from Spain, Peru, India, and Japan. A tfs3 remnant (6 kb) was found in an archived stock of reference strain J99, although it was not included in this strain's published genome sequence. PCR and DNA sequence analyses indicated the following. (i) tfs3's ends are conserved. (ii) Right-end insertion occurred at one specific site in a chromosomal region that is varied in gene content and arrangement, the "plasticity zone." (iii) Left-end insertion occurred at different sites in each of nine strains studied. (iv) Sequences next to the right-end target in tfs3-free strains were absent from most strains carrying full-length tfs3 elements. These patterns suggested insertion by a transposition-like event, but one in which targets are chosen with little or no specificity at the left end and high specificity at the right end, thereby deleting the intervening DNA.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Campus Box 8230, Washington University School of Medicine, St. Louis, MO 63110. Phone: (314) 362-2772. Fax: (314) 362-1232. E-mail: berg{at}borcim.wustl.edu.

{dagger} Present address: National Institute of Cholera and Enteric Diseases, Scheme-XM, Beliaghata, Calcutta-700010, India.

Journal of Bacteriology, July 2003, p. 3764-3772, Vol. 185, No. 13
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.13.3764-3772.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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