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Infection and Immunity, December 2002, p. 6726-6733, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6726-2733.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Chromosomal DNA Deletions Explain Phenotypic Characteristics of Two Antigenic Variants, Phase II and RSA 514 (Crazy), of the Coxiella burnetii Nine Mile Strain{dagger}

T. A. Hoover,1 D. W. Culp,2 M. H. Vodkin,3 J. C. Williams,4 and H. A. Thompson5,6*

Bacteriology Division, USAMRIID, Ft. Detrick, Frederick, Maryland 21701,1 Division of Laboratories, Illinois Department of Public Health, Springfield, Illinois 62794,2 Center for Economic Entomology, Illinois Natural History Survey, Champaign, Illinois 61821,3 Wyeth Biopharma, Andover, Massachusetts 01810,4 Viral and Rickettsial Zoonoses Branch, National Center for Infectious Diseases, Centers for Disease Control, Atlanta, Georgia 30333,5 Department of Microbiology and Immunology, R. E. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 265066

Received 18 April 2002/ Returned for modification 30 May 2002/ Accepted 20 August 2002

After repeated passages through embyronated eggs, the Nine Mile strain of Coxiella burnetii exhibits antigenic variation, a loss of virulence characteristics, and transition to a truncated lipopolysaccharide (LPS) structure. In two independently derived strains, Nine Mile phase II and RSA 514, these phenotypic changes were accompanied by a large chromosomal deletion (M. H. Vodkin and J. C. Williams, J. Gen. Microbiol. 132:2587-2594, 1986). In the work reported here, additional screening of a cosmid bank prepared from the wild-type strain was used to map the deletion termini of both mutant strains and to accumulate all the segments of DNA that comprise the two deletions. The corresponding DNAs were then sequenced and annotated. The Nine Mile phase II deletion was completely nested within the deletion of the RSA 514 strain. Basic alignment and homology studies indicated that a large group of LPS biosynthetic genes, arranged in an apparent O-antigen cluster, was deleted in both variants. Database homologies identified, in particular, mannose pathway genes and genes encoding sugar methylases and nucleotide sugar epimerase-dehydratase proteins. Candidate genes for addition of sugar units to the core oligosaccharide for synthesis of the rare sugar 6-deoxy-3-C-methylgulose (virenose) were identified in the deleted region. Repeats, redundancies, paralogous genes, and two regions with reduced G+C contents were found within the deletions.

* Corresponding author. Mailing address: Viral and Rickettsial Zoonoses Branch, National Center for Infectious Diseases, Centers for Disease Control, Mailstop G-13, 1600 Clifton Road NE, Atlanta, GA 30333. Phone: (404) 639-1083. Fax: (404) 639-1056. E-mail: hct2{at}cdc.gov. {dagger} We dedicate this article to the memory of Paul Fiset, who, along with M. Stoker, first described antigenic phase variation in Coxiella.

Editor: J. T. Barbieri

Infection and Immunity, December 2002, p. 6726-6733, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6726-2733.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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