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Infection and Immunity, March 2000, p. 1574-1586, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Molecular and Evolutionary Characterization of the cp32/18 Family
of Supercoiled Plasmids in Borrelia burgdorferi
297
Melissa J.
Caimano,1
Xiaofeng
Yang,2
Taissia G.
Popova,2
Michael L.
Clawson,1
Darrin R.
Akins,3
Michael V.
Norgard,2 and
Justin D.
Radolf1,4,5,*
Center for Microbial
Pathogenesis1 and Departments of
Medicine4 and
Microbiology,5 University of Connecticut
Health Center, Farmington, Connecticut 06030; Department of
Microbiology, University of Texas Southwestern Medical Center,
Dallas, Texas 75235;2 and Department of
Microbiology and Immunology, University of Oklahoma Health Sciences
Center, Oklahoma City, Oklahoma 731903
Received 20 September 1999/Returned for modification 17 November
1999/Accepted 26 November 1999
In this study, we characterized seven members of the cp32/18 family
of supercoiled plasmids in Borrelia burgdorferi
297. Complete sequence analysis of a 21-kb plasmid (cp18-2) confirmed
that the strain 297 plasmids are similar in overall content and
organization to their B31 counterparts. Of the 31 open reading frames
(ORFs) in cp18-2, only three showed sequence relatedness to proteins with known functions, and only one, a ParA/SopA ortholog, was related
to nonborrelial polypeptides. Besides the lipoproteins, none of the
ORFs appeared likely to encode a surface-exposed protein. Comparison
with the B31 genomic sequence indicated that paralogs for most of the
ORFs in cp18-2 can be identified on other genetic elements. cp18-2 was
found to lack a 9- to 10-kb fragment present in the 32-kb homologs
which, by extrapolation from the B31 cp32 sequences, contains at least
15 genes presumed to be unnecessary for plasmid maintenance. Sequence
analysis of the lipoprotein-encoding variable loci provided evidence
that recombinatorial processes within these regions may result in the
acquisition of exogenous DNA. Pairwise analysis with random shuffling
revealed that the multiple lipoproteins (Mlp; formerly designated 2.9 LPs) fall into two distinct homology groups which appear to have arisen by gene fusion events similar to those recently proposed to have generated the three OspE, OspF, and Elp lipoprotein families (D. R. Akins, M. J. Caimano, X. Yang, F. Cerna, M. V. Norgard,
and J. D. Radolf, Infect. Immun. 67:1526-1532, 1999). Comparative analysis of the variable regions also indicated that recombination within the loci of each plasmid may occur independently. Last, comparison of variable loci revealed that the cp32/18 plasmid complements of the B31 and 297 isolates differ substantially, indicating that the two strains have been subject to divergent adaptive
pressures. In addition to providing evidence for two different types of
recombinatorial events involving cp32/18 plasmids, these findings
underscore the need for genetic analysis of diverse borrelial isolates
in order to elucidate the Lyme disease spirochete's complex parasitic strategies.
*
Corresponding author. Mailing address: Center for
Microbial Pathogenesis, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030-3710. Phone: (860) 679-8129. Fax:
(860) 679-8130. E-mail: JRadolf{at}up.uchc.edu.
Infection and Immunity, March 2000, p. 1574-1586, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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