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Journal of Bacteriology, November 1999, p. 6779-6787, Vol. 181, No. 21
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Vibrio cholerae O139 Calcutta Bacteriophage CTXphi Is Infectious and Encodes a Novel Repressor

Brigid M. Davis, Harvey H. Kimsey, William Chang, and Matthew K. Waldor*

Tufts University School of Medicine and Division of Geographic Medicine and Infectious Diseases, Tupper Research Institute, Boston, Massachusetts 02111

Received 11 May 1999/Accepted 30 August 1999

CTXphi is a lysogenic, filamentous bacteriophage. Its genome includes the genes encoding cholera toxin (ctxAB), one of the principal virulence factors of Vibrio cholerae; consequently, nonpathogenic strains of V. cholerae can be converted into toxigenic strains by CTXphi infection. O139 Calcutta strains of V. cholerae, which were linked to cholera outbreaks in Calcutta, India, in 1996, are novel pathogenic strains that carry two distinct CTX prophages integrated in tandem: CTXET, the prophage previously characterized within El Tor strains, and a new CTX Calcutta prophage (CTXcalc). We found that the CTXcalc prophage gives rise to infectious virions; thus, CTXETphi is no longer the only known vector for transmission of ctxAB. The most functionally significant differences between the nucleotide sequences of CTXcalcphi and CTXETphi are located within the phages' repressor genes (rstRcalc and rstRET, respectively) and their RstR operators. RstRcalc is a novel, allele-specific repressor that regulates replication of CTXcalcphi by inhibiting the activity of the rstAcalc promoter. RstRcalc has no inhibitory effect upon the classical and El Tor rstA promoters, which are instead regulated by their cognate RstRs. Consequently, production of RstRcalc renders a CTXcalc lysogen immune to superinfection by CTXcalcphi but susceptible (heteroimmune) to infection by CTXETphi . Analysis of the prophage arrays generated by sequentially integrated CTX phages revealed that pathogenic V. cholerae O139 Calcutta probably arose via infection of an O139 CTXETphi lysogen by CTXcalcphi .

* Corresponding author. Mailing address: Tufts University School of Medicine and Division of Geographic Medicine and Infectious Diseases, Tupper Research Institute, Tufts-New England Medical Center 041, 750 Washington St., Boston, MA 02111. Phone: (617) 636-7618. Fax: (617) 636-5292. E-mail: matthew.waldor{at}es.nemc.org.

Journal of Bacteriology, November 1999, p. 6779-6787, Vol. 181, No. 21
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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