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Infect Immun, July 1998, p. 3088-3094, Vol. 66, No. 7
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Heterogeneity in Levels of Vacuolating Cytotoxin Gene (vacA) Transcription among Helicobacter pylori Strains

M. H. Forsyth,1 J. C. Atherton,2 M. J. Blaser,1,3 and T. L. Cover1,3,*

Departments of Medicine and Microbiology and Immunology, Vanderbilt University School of Medicine,1 and Veterans Affairs Medical Center,3 Nashville, Tennessee, and Division of Gastroenterology and Institute of Infections and Immunity, University of Nottingham, Nottingham, United Kingdom2

Received 2 September 1997/Returned for modification 4 November 1997/Accepted 24 April 1998

Broth culture supernatants from Tox+ Helicobacter pylori strains induce vacuolation of HeLa cells in vitro and contain VacA in concentrations that are higher than those found in supernatants from Tox- H. pylori strains. To investigate the basis for this phenomenon, we analyzed the transcription of the vacuolating cytotoxin gene (vacA) in eight Tox+ strains (each with a type s1/m1 vacA genotype) and nine Tox- strains (each with a type s2/m2 vacA genotype). Most of the Tox+ and Tox- strains tested used the same vacA transcriptional start point, but Tox+ strains yielded significantly stronger primer extension signal intensities than did Tox- strains (mean densitometry values of 15.8 ± 1.9 versus 8.9 ± 1.7, P = 0.0016). Correspondingly, when we introduced vacA::xylE transcriptional fusions into the chromosomes of a Tox+ strain (60190) and a Tox- strain (86-313), the level of XylE activity in 60190 vacA::xylE was about 30-fold higher than that in 86-313 vacA::xylE. Sequence analysis and promoter exchange experiments indicated that the different levels of vacA transcription in these two strains cannot be explained solely by a difference in promoter strength. These data indicate that Tox+ and Tox- H. pylori strains typically differ not only in the VacA amino acid sequence but also in the level of vacA transcription.

* Corresponding author. Mailing address: Division of Infectious Diseases, Medical Center North A3310, Vanderbilt University School of Medicine, Nashville, TN 37232-2605. Phone: (615) 322-2035 Fax: (615) 343-6160 E-mail: COVERTL{at}ctrvax.vanderbilt.edu.

Infect Immun, July 1998, p. 3088-3094, Vol. 66, No. 7
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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