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Journal of Bacteriology, November 1998, p. 5567-5573, Vol. 180, No. 21
Department of Microbiology, Colorado State
University, Fort Collins, Colorado 80523-1677
Received 12 May 1998/Accepted 18 August 1998
The Mycobacterium avium complex is a source of
disseminated infections in patients with advanced AIDS. This group of
mycobacteria is distinguished by the presence of highly antigenic,
surface-exposed glycopeptidolipids, and these glycolipids possess
variant oligosaccharide structures that are the chemical basis of the
28 distinct serovars of the M. avium complex. We previously
described the ser2 gene cluster, encoding the synthesis of
the haptenic oligosaccharide (2,3-dimethylfucose-rhamnose-6-deoxytalose-) of the serovar 2-specific glycopeptidolipid, and revealed a locus (ser2A) encoding a
putative rhamnosyltransferase. Sequencing of the ser2A
locus demonstrated the presence of three open reading frames, two of
which yielded significant homology to several glycosyltransferases, and
the deduced amino acid sequences of these two putative
glycosyltransferases had 63% identity. These two genes were expressed
in Mycobacterium smegmatis, and the resulting recombinant
glycopeptidolipids were characterized by thin-layer chromatography and
gas chromatography-mass spectrometry. These analyses demonstrated that
only one of these genes, termed rtfA, encoded the
rhamnosyltransferase responsible for the transfer of rhamnose to
6-deoxytalose. The identification of rtfA will permit
further evaluation of glycopeptidolipid biosynthesis and the
construction of isogenic mutants of multiple M. avium complex serovars. Moreover, such mutants will help define the role of
glycopeptidolipids in the intracellular survival of these bacteria.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Identification and Recombinant Expression of a
Mycobacterium avium Rhamnosyltransferase Gene
(rtfA) Involved in Glycopeptidolipid Biosynthesis
*
Corresponding author. Mailing address: Department of
Microbiology, Colorado State University, Fort Collins, CO 80523-1677. Phone: (970) 491-6549. Fax: (970) 491-1815. E-mail:
jbelisle{at}cvmbs.colostate.edu.
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