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Journal of Bacteriology, October 2003, p. 5822-5830, Vol. 185, No. 19
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.19.5822-5830.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Complete Sequence of Virulence Plasmid pJM1 from the Marine Fish Pathogen Vibrio anguillarum Strain 775

Manuela Di Lorenzo,1 Michiel Stork,1 Marcelo E. Tolmasky,2 Luis A. Actis,3 David Farrell,4 Timothy J. Welch,5 Lidia M. Crosa,1 Anne M. Wertheimer,1,6 Qian Chen,7 Patricia Salinas,8 Lillian Waldbeser,9 and Jorge H. Crosa1*

Department of Molecular Microbiology and Immunology,1 Department of Pathology,4 Department of Medicine, Oregon HealthScience University, Portland, Oregon 97201,6 Department of Biological Science, California State University Fullerton, Fullerton, California 92834-6850,2 Department of Microbiology, Miami University, Oxford, Ohio 45056,3 National Center for Water Aquaculture, Agricultural Research Service/U.S. Department of Agriculture, Kearneysville, West Virginia 25430,5 Eisai Research Institute, Wilmington, Massachusetts 01887,7 Department of Biological Sciences, Imperial College of Science, Technology and Medicine, London, United Kingdom,8 Department of Microbiology, Texas A&M University, Corpus Christi, Texas 784129

Received 19 May 2003/ Accepted 3 July 2003

The virulence plasmid pJM1 enables the fish pathogen Vibrio anguillarum, a gram-negative polarly flagellated comma-shaped rod bacterium, to cause a highly fatal hemorrhagic septicemic disease in salmonids and other fishes, leading to epizootics throughout the world. The pJM1 plasmid 65,009-nucleotide sequence, with an overall G+C content of 42.6%, revealed genes and open reading frames (ORFs) encoding iron transporters, nonribosomal peptide enzymes, and other proteins essential for the biosynthesis of the siderophore anguibactin. Of the 59 ORFs, approximately 32% were related to iron metabolic functions. The plasmid pJM1 confers on V. anguillarum the ability to take up ferric iron as a complex with anguibactin from a medium in which iron is chelated by transferrin, ethylenediamine-di(o-hydroxyphenyl-acetic acid), or other iron-chelating compounds. The fatDCBA-angRT operon as well as other downstream biosynthetic genes is bracketed by the homologous ISV-A1 and ISV-A2 insertion sequences. Other clusters on the plasmid also show an insertion element-flanked organization, including ORFs homologous to genes involved in the biosynthesis of 2,3-dihydroxybenzoic acid. Homologues of replication and partition genes are also identified on pJM1 adjacent to this region. ORFs with no known function represent approximately 30% of the pJM1 sequence. The insertion sequence elements in the composite transposon-like structures, corroborated by the G+C content of the pJM1 sequence, suggest a modular composition of plasmid pJM1, biased towards acquisition of modules containing genes related to iron metabolic functions. We also show that there is considerable microheterogeneity in pJM1-like plasmids from virulent strains of V. anguillarum isolated from different geographical sources.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, L220, Oregon Health and Science University, 3181 S.W. Sam Jackson Park Rd., Portland, OR 97201. Phone: (503) 494-7583. Fax: (503) 494-6862. E-mail: crosajor{at}ohsu.edu.

Journal of Bacteriology, October 2003, p. 5822-5830, Vol. 185, No. 19
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.19.5822-5830.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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