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Antimicrobial Agents and Chemotherapy, October 2001, p. 2971-2972, Vol. 45, No. 10
0066-4804/01/$04.00+0 DOI: 10.1128/AAC.45.10.2971-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
LETTERS TO THE EDITOR
Plasmid-Mediated Rifampin Resistance Encoded by an
arr-2-Like Gene Cassette in Klebsiella
pneumoniae Producing an ACC-1 Class C 
-Lactamase
 |
LETTER |
For Pseudomonas spp., two plasmid-mediated mechanisms
of resistance to rifampin have been reported: one involves an efflux pump (1), and the other involves an ADP-ribosylating
transferase (3) encoded by a gene (arr-2) that
is integrated as a gene cassette in a class 1 integron
(10). Recently, plasmid-mediated rifampin resistance
(Arr-2) was described for various Enterobacteriaceae from
Southeast Asia producing the extended-spectrum 
-lactamase VEB-1
(4, 6). The genes encoding these mechanisms of resistance are integron located as gene cassettes (4, 6).
We recently described an outbreak in France of Klebsiella
pneumoniae isolates that produce the ACC-1 -lactamase and that are resistant to rifampin, the first of which (strain SLK54) was isolated from a patient previously hospitalized in Tunisia
(7). Resistance to ceftazidime and rifampin was
cotransferred by conjugation to Escherichia coli K-12 strain
HB101 for all strains (rifampin MICs, >256 µg
ml
1) except one (KP SLK55) (rifampin MIC, 16 µg
ml1). Plasmid DNA from the transconjugant E. coli HB101 of the first isolate in the outbreak
(K. pneumoniae SLK54) was partially digested with
Sau3A (Roche Molecular Biochemicals, Meylan, France) and ligated (T4 DNA ligase; Amersham Pharmacia Biotech, Saclay,
France) into the BamHI site of the cloning vector
pBK-CMV Kanr (Stratagene, La Jolla, Calif.)
(8). The recombinant plasmid, pRIF-1, which harbored the
smallest insert (2 kb) was sequenced (9). Analysis of the
DNA sequence revealed an open reading frame of 453 nucleotides (Fig.
1) predicting a 150-amino-acid sequence.
This protein was 99% identical to the rifampin ADP-ribosylating transferase Arr-2. The predicted rifampin ADP-ribosylating transferase differed from Arr-2 by a Lys98Arg substitution. In Pseudomonas aeruginosa the location of the gene encoding this protein is
unclear (10), although in Enterobacteriaceae it
has been found to be plasmid located (4, 6). However, in
both cases, the arr-2 gene was a gene cassette located in a
class 1 integron. In K. pneumoniae SLK54, the
plasmid-mediated arr-2-like gene was also a gene cassette,
located immediately downstream of the integrase and the promoter region
(Fig. 1). The expression of the arr-2-like gene is probably
driven by the Pant promoter and not the P2 promoter because
the insertion of three G residues increasing the spacing between the
35 and 10 boxes to 17 was absent in the P2 promoter (Fig. 1)
(2). The configuration of the Pant promoter
was a hybrid combining the 35 box (TGGACA), found in weak
Pant promoters, and the 10 box (TAAACT), found in strong
Pant promoters, and has been shown to have
intermediate strength (5). The 5' coding sequence
of the integron, corresponding to the last 214 bp (71 amino acids) of
the integrase gene, was truncated by the insertion of the IS26 sequence
and resulted in a nonfunctional integrase. A similar observation was
made for In53, which carries the arr-2 gene cassette in
E. coli (6), in which the IS26 truncated the integrase gene and especially the Pant promoter.
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FIG. 1.
Nucleotide sequence of the arr-2 gene and its
genetic environment. The 5' and 3' ends, the 35 and 10 promoter
regions, the core sites and inverse core sites, and the 59-bp element
are indicated.
|
|
The rifampin ADP-ribosylating transferase Arr-2 was previously detected
in strains of P. aeruginosa and
Enterobacteriaceae isolated from patients in Southeast Asia.
These strains also produce the VEB-1 extended-spectrum -lactamase
(4, 6, 10). We now report the plasmid-mediated rifampin
resistance encoded by the arr-2-like gene cassette in
K. pneumoniae which also produces the plasmid-mediated
cephalosporinase ACC-1 and which was isolated in France from a patient
previously hospitalized in Tunisia (7).
The EMBL accession number for the nucleotide sequence reported in this
paper is AJ277027.
| |
FOOTNOTES |
*
Phone: 33 1 56 01 70 18 Fax: 33 1 56 01 61 08 E-mail:
guillaume.arlet{at}tnn.ap-hop-paris.fr
| |
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| | | | |
Guillaume Arlet*
David Nadjar
Service de
Bactériologie
Hôpital Tenon
UFR Saint Antoine
4
rue de la Chine
75970 Paris Cedex 20, France
|
| | | | |
Jean-Louis Herrmann
Jean-Luc Donay
Martine Rouveau
Philippe H. Lagrange
Service de
Microbiologie
Hôpital Saint-Louis
UFR Lariboisière
Saint-Louis
Paris, France
|
| | | | |
Alain Philippon
Service de Microbiologie
Hôpital
Cochin
Paris, France
|
Antimicrobial Agents and Chemotherapy, October 2001, p. 2971-2972, Vol. 45, No. 10
0066-4804/01/$04.00+0 DOI: 10.1128/AAC.45.10.2971-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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