[en] The location and environment of the acquired blaIMP gene, which encodes the IMP-1 metallo-beta-lactamase, were investigated in a Japanese Pseudomonas aeruginosa clinical isolate (isolate 101/1477) that produced the enzyme. In this isolate, blaIMP was carried on a 36-kb plasmid, and similar to the identical alleles found in Serratia marcescens and Klebsiella pneumoniae clinical isolates, it was located on a mobile gene cassette inserted into an integron. The entire structure of this integron, named In31, was determined. In31 is a class 1 element belonging to the same group of defective transposon derivatives that originated from Tn402-like ancestors such as In0, In2, and In5. The general structure of In31 appeared to be most closely related to that of In5 from pSCH884, suggesting a recent common phylogeny for these two elements. In In31, the blaIMP cassette is the first of an array of five gene cassettes that also includes an aacA4 cassette and three original cassettes that have never been described in other integrons. The novel cassettes carry, respectively, (i) a new chloramphenicol acetyltransferase-encoding allele of the catB family, (ii) a qac allele encoding a new member of the small multidrug resistance family of proteins, and (iii) an open reading frame encoding a protein of unknown function. All the resistance genes carried on cassettes inserted in In31 were found to be functional in decreasing the in vitro susceptibilities of host strains to the corresponding antimicrobial agents.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Laraki, Nadine ; Université de Liège - ULiège > Intervention et gestion en activités physiques et sportives
Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Thamm, Iris ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Riccio, M. L.
Amicosante, G.
Frère, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Rossolini, G. M.
Language :
English
Title :
Structure of In31, a Blaimp-Containing Pseudomonas Aeruginosa Integron Phyletically Related to In5, Which Carries an Unusual Array of Gene Cassettes
Publication date :
April 1999
Journal title :
Antimicrobial Agents and Chemotherapy
ISSN :
0066-4804
eISSN :
1098-6596
Publisher :
American Society for Microbiology, Washington, United States - District of Columbia
Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment search tool. J. Mol. Biol. 215:403-410.
Arakawa, Y., M. Murakami, K. Suzuki, H. Ito, R. Wacharotayankun, S. Ohsuka, N. Kato, and M. Ohta. 1995. A novel integron-like element carrying the metallo-β-lactamase gene blaIMP. Antimicrob. Agents Chemother. 39: 1612-1615.
Bissonnette, L., and P. H. Roy. 1992. Characterization of In0 of Pseudomonas aeruginosa plasmid pVS1, an ancestor of integrons of multiresistance plasmids and transposons of gram-negative bacteria. J. Bacteriol. 174:1248-1257.
Bradford, M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254.
Brown, H. J., H. W. Stokes, and R. M. Hall. 1996. The integrons In0, In2, and In5 are defective transposon derivatives. J. Bacteriol. 178:4429-4437.
Bunny, K. L., R. M. Hall, and H. W. Stokes. 1995. New mobile gene cassettes containing an aminoglycoside resistance gene, aacA7, and a chloramphenicol resistance gene, catB3, in an integron in pBWH301. Antimicrob. Agents Chemother. 39:686-693.
Collis, C. M., and R. M. Hall. 1992. Site-specific deletion and rearrangement of integron insert genes catalyzed by the integron DNA integrase. J. Bacteriol. 174:1574-1585.
Collis, C. M., and R. M. Hall. 1992. Gene cassettes from the insert region of integrons are excised as covalently closed circles. Mol. Microbiol. 6:2875-2885.
Collis, C. M., G. Grammaticopoulos, J. Briton, H. W. Stokes, and K. M. Hall. 1993. Site-specific insertion of gene cassettes into integrons. Mol. Microbiol. 9:41-52.
Collis, C. M., and R. M. Hall. 1995. Expression of antibiotic resistance genes in the integrated cassettes of integrons. Antimicrob. Agents Chemother. 39:155-162.
Devereux, J., P. Haeberli, and O. Smithies. 1984. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 12:387-395.
Galimand, M., T. Lambert, G. Gerbaud, and P. Couvalin. 1993. Characterization of the aac(6′)-lb gene encoding an aminoglycoside 6′-N-acetyltransferase in Pseudomonas aeruginosa BM2656. Antimicrob. Agents Chemother. 37:1456-1462.
Glaser, P., F. Kunst, M. Arnaud, M. P. Coudart, W. Gonzales, M. F. Hullo, M. Ionescu, B. Lubochinsky, L. Marcelino, I. Moszer, E. Presecan, M. Santana, E. Schneider, J. Schweizer, A. Vertes, G. Rapoport, and A. Danchin. 1993. Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325 degrees to 333 degrees. Mol. Microbiol. 10:371-384.
Goldberg, J. B., and D. E. Ohman. 1984. Cloning and expression in Pseudomonas aeruginosa of a gene involved in the production of alginate. J. Bacteriol. 158:1115-1121.
Grantham, K., C. Gautier, M. Gouy, M. Jacobzone, and R. Mercier. 1981. Codon catalog usage is a genome strategy modulated for gene expressivity. Nucleic Acids Res. 9:43-74.
Guerineau, F., L. Brooks, and P. Mullineaux. 1990. Expression of sulphonamide resistance gene from plasmid R46. Plasmid 23:35-41.
Hall, R. M., and C. M. Collis. 1995. Mobile gene cassettes and integrons: capture and spread of genes by site-specific recombination. Mol. Microbiol. 15:593-600.
Hall, R. M., and C. Vockler. 1987. The region of the IncN plasmid R46 coding for resistance to β-lactam antibiotics, streptomycin/spectinomycin and sulphonamides is closely related to antibiotic resistance segments found in IncW plasmids and in Tn21-like transposons Nucleic Acids Res. 15:7491-7501.
Hall, R. M., D. E. Brookes, and H. W. Stokes. 1991. Site-specific insertion of genes into integrons: role of the 59-base element and determination of the recombination cross-over point. Mol. Microbiol. 5:1941-1959.
Hall, R. M., H. J. Brown, D. E. Brookes, and H. W. Stokes. 1994. Integrons found in different locations have identical 5′ ends but variable 3′ ends. J. Bacteriol. 176:6286-6294.
Hirakata, Y., K. Izumikawa, T. Yamaguchi, H. Takemura, H. Tanaka, R. Yoshida, J. Matsuda, M. Nakano, K. Tomono, S. Maesaki, M. Kaku, Y. Yamada, S. Kamihira, and S. Kohno. 1998. Rapid detection and evaluation of clinical characteristics of emerging multiple-drug-resistant gram-negative rods carrying the metallo-β-lactamase gene blaIMP. Antimicrob. Agents Chemother. 42:2006-2011.
Ito, H., Y. Arakawa, S. Ohsuka, R. Wacharotayankun, N. Kato, and M. Ohta. 1995. Plasmid-mediated dissemination of the metallo-β-lactamase gene blaIMP, among clinically isolated strains of Serratia marcescens. Antimicrob. Agents Chemother. 39:824-829.
Iyobe, S., H. Yamada, and S. Minami. 1996. Insertion of a carbapenemase gene cassette into an integron of a Pseudomonas aeruginosa plasmid. J. Antimicrob. Chemother. 38:1114-1115.
Kholodii, G. Y., S. Z. Mindlin, I. A. Bass, O. V. Yurieva, S. V. Minakhina, and V. G. Nikiforov. 1995. Four genes, two ends, and a res region are involved in transposition on Tn5053: a paradigm for a novel family of transposons carrying either a mer operon or an integron. Mol. Microbiol. 17:1189-1200.
Kupzig, S., and P. M. Bennett. 1994. EMBL/GenBank database entry X82455.
Laraki, N., N. Franceschini, G. M. Rossolini, P. Santucci, C. Meunier, E. de Pauw, G. Amicosante, J.-M. Frére, and M. Galleni. 1999. Biochemical characterization of the Pseudomonas aeruginosa 101/1477 metallo-β-lactamase IMP-1 produced by Escherichia coli. Antimicrob. Agents Chemother. 43:813-817.
Laraki, N., M. Galleni, G. M. Rossolini, and J.-M. Frére. Unpublished results.
Lévesque, C., S. Brassard, J. Lapointe, and P. H. Roy. 1994. Diversity and relative strength of tandem promoters for the antibiotic resistance genes of several integrons. Gene 142:49-54.
Littlejohn, T. G., D. DiBerardino, L. J. Messerotti, S. J. Spiers, and R. A. Skurray. 1991. Structure and evolution of a family of genes encoding antiseptic and disinfectant resistance in Staphylococcus aureus. Gene 101:59-66.
Mabilat, C., J. Lourençao-Vital, S. Goussard, and P. Courvalin. 1992. A new example of physical linkage between Tn1 and Tn21: the antibiotic multiple-resistance region of plasmid pCFF04 encoding extended-spectrum β-lactamase TEM-3. Mol. Gen. Genet. 235:113-121.
Martinez, E., and F. de la Cruz. 1990. Genetic elements involved in Tn21 site-specific integration, a novel mechanism for the dissemination of antibiotic resistance genes. EMBO J. 9:1275-1281.
Marumo, K., A. Takeda, Y. Nakamura, and K. Nakaya. 1995. Purification and characterization of metallo-β-lactamase from Serratia marcescens. Microbiol. Immunol. 39:27-33.
Mugnier, P., I. Podglajen, F. W. Goldstein, and E. Collatz. 1998. Carbapenems as inhibitors of OXA-13, a novel, integron-encoded β-lactamase in Pseudomonas aeruginosa. Microbiology 144:1021-1031.
National Committee for Clinical Laboratory Standards. 1997. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. 4th ed. Approved standard. NCCLS document M7-A4. National Committee for Clinical Laboratory Standards, Wayne, Pa.
Nobuta, K., M. E. Tolmasky, L. M. Crosa, and J. H. Crosa. 1988. Sequencing and expression of the 6′-N-acetyltransferase gene of transposon Tn1331 from Klebsiella pneumoniae. J. Bacteriol. 170:3769-3773.
Osano, E., Y. Arakawa, R. Wacharotayankun, M. Ohta, T. Horii, H. Ito, F. Yoshimura, and N. Kato. 1994. Molecular characterization of an enterobacterial metallo-β-lactamase found in a clinical isolate of Serratia marcescens that shows imipenem resistance. Antimicrob. Agents Chemother. 38:71-78.
Parent, R., and P. H. Roy. 1992. The chloramphenicol acetyl-transferase gene of Tn2424: a new breed of cat. J. Bacteriol. 174:2891-2897.
Paulsen, I. T., R. A. Skurray, R. Tam, M. H. Saier, Jr., R. J. Turner, J. H. Weiner, E. B. Goldberg, and L. L. Grinius. 1996. The SMR family: a novel family of multidrug efflux proteins involved with the efflux of lipophilic drugs. Mol. Microbiol. 19:1167-1175.
Paulsen, I. T., T. G. Littlejohn, P. Rådström, L. Sundström, O. Sköld, G. Swedberg, and R. A. Skurray. 1993. The 3′ conserved segment of integrons contains a gene associated with multidrug resistance to antiseptics and disinfectants. Antimicrob. Agents Chemother. 37:761-768.
Payne, D. J. 1993. Metallo-β-lactamases - a new therapeutic challenge. J. Med. Microbiol. 39:93-99.
Pearson, W. R. 1990. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol. 183:63-98.
Ploy, M.-C., P. Courvalin, and T. Lambert. 1998. Characterization of In40 of Enterobacter aerogenes BM2688, a class 1 integron with two new gene cassettes, cm1A2 and qacF. Antimicrob. Agents Chemother. 42:2557-2563.
Preston, K. E., M. A. Kacica, R. J. Limberger, W. A. Archinal, and R. A. Venezia. 1997. The resistance and integrase genes of pACM1, a conjugative multiple-resistance plasmid, from Klebsiella oxytoca. Plasmid 37:105-118.
Pridmore, R. D. 1987. New and versatile cloning vectors with kanamycin resistance marker. Gene 56:309-312.
Purewal, A. S. 1991. Nucleotide sequence of the ethidium efflux gene from Escherichia coli. FEMS Microbiol. Lett. 66:229-231.
Rådström, P., O. Sköld, G. Swedberg, J. Flensburg, P. H. Roy, and L. Sundström. 1994. Transposon Tn5090 of plasmid R751, which carries an integron, is related to Tn7, Mu, and the retroelements. J. Bacteriol. 176: 3257-3268.
Rasmussen, B. A., and K. Bush. 1997. Carbapenem-hydrolyzing β-lactamases. Antimicrob. Agents Chemother. 41:223-232.
Recchia, G. D., and R. M. Hall. 1995. Gene cassettes: a new class of mobile element. Microbiology 141:3015-3027.
Reese, M. G. 18 December 1998, revision date. Promoter prediction by neural network program. [Online.] Drosophila Genome Center, Lawrence Berkeley National Laboratory, Berkeley, Calif. http://www-hgc.lbl.gov./ projects/promoter.html. [20 December 1998, last date accessed.]
Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual, 2nd cd. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y.
Sanger, F., S. Nicklen, and A. R. Coulson. 1977. DNA sequencing with chain terminating inhibitors. Proc. Natl. Acad. Sci. USA 74:5463-5467.
Senda, K., Y. Arakawa, K. Nakashima, H. Ito, S. Ichiyama, K. Shimokata, N. Kato, and M. Ohta. 1996. Multifocal outbreaks of metallo-β-lactamase-producing Pseudomonas aeruginosa resistant to broad-spectrum β-lactams, including carbapenems. Antimicrob. Agents Chemother. 40:349-353.
Senda, K., Y. Arakawa, S. Ichiyama, K. Nakashima, H. Ito, S. Ohsuka, K. Shimokata, N. Kato, and M. Ohta. 1996. PCR detection of metallo-β-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum β-lactams. J. Clin. Microbiol. 34:2909-2913.
Shaw, K. J., P. N. Rather, R. S. Hare, and G. H. Miller. 1993. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol. Rev. 57:138-163.
Stokes, H. W., and R. M. Hall. 1989. A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons. Mol. Microbiol. 3:1669-1683.
Stokes, H. W., D. B. O'Gorman, G. D. Recchia, M. Parsekhian, and R. M. Hall. 1997. Structure and function of 59-base element recombination sites associated with mobile gene cassettes. Mol. Microbiol. 26:731-745.
Tennigkeit, J., and H. Matzura. 1991. Nucleotide sequence analysis of a chloramphenicol-resistance determinant from Agrobacterium tumefaciens and identification of its gene product. Gene 98:113-116.
Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22:4673-4680.
Toriya, M., M. Sakakibara, K. Matsushita, and T. Morohoshi. 1992. Nucleotide sequence of aminoglycoside 6′-N-acetyltransferase [AAC(6′)] determinant from Serratia sp. 45. Chem. Pharm. Bull. 40:2473-2477.
Vézina, G., and R. C. Levesque. 1991. Molecular characterization of the class II multiresistance transposable element in Tn1043 from Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 35:313-321.
Watanabe, M., S. Iyobe, M. Inoue, and S. Mitsuhashi. 1991. Transferable imipenem resistance in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 35:147-151.
Yamaguchi, H., M. Nukaya, and T. Sawai. 1994. EMBL/GenBank database entry D29636.
