Tn2009, a Tn916-Like Element Containing mef(E) in Streptococcus pneumoniae

ABSTRACT The association between the macrolide efflux gene mef(E) and the tet(M) gene was studied in two clinical strains of Streptococcus pneumoniae that belonged to serotypes 19F and 6A, respectively, and that were resistant to both tetracycline and erythromycin. The mef(E)-carrying element mega (macrolide efflux genetic assembly; 5,511 bp) was found to be inserted into a Tn916-like genetic element present in the chromosomes of the two pneumococcal strains. In both strains, mega was integrated at the same site, an open reading frame identical to orf6 of Tn916. The new composite element, Tn2009, was about 23.5 kb and, with the exception of the tet(M)-coding sequence, appeared to be identical in both strains. By sequencing of the junction fragments of Tn2009 at the site of insertion into the chromosome, it was possible to show that (i) the insertion site was identical in the two clinical strains and (ii) the integration of Tn2009 caused a 9.5 kb-deletion in the pneumococcal chromosome. It was not possible to detect the conjugal transfer of Tn2009 to a recipient pneumococcal strain; however, transfer of the whole element by transformation was shown to occur. It is possible to hypothesize that Tn2009 relies on transformation for its spread among clinical strains of S. pneumoniae.

[1]  G. Pozzi,et al.  Method for introducing specific and unmarked mutations into the chromosome of Streptococcus pneumoniae , 2004, Molecular biotechnology.

[2]  N. Shoemaker,et al.  Destruction of low efficiency markers is a slow process occurring at a heteroduplex stage of transformation , 2004, Molecular and General Genetics MGG.

[3]  D. George,et al.  Slow excision repair in an mfd mutant of Escherichia coli B/r , 2004, Molecular and General Genetics MGG.

[4]  P. Huovinen,et al.  Antimicrobial susceptibility patterns and macrolide resistance genes of viridans group streptococci from normal flora. , 2003, The Journal of antimicrobial chemotherapy.

[5]  M. Roberts,et al.  Presence of the tet(O) Gene in Erythromycin- and Tetracycline-Resistant Strains of Streptococcus pyogenes and Linkage with either the mef(A) or the erm(A) Gene , 2003, Antimicrobial Agents and Chemotherapy.

[6]  A. Pantosti,et al.  Phenotypic and Genotypic Characterization of Two Penicillin-Susceptible Serotype 6B Streptococcus pneumoniae Clones Circulating in Italy , 2003, Journal of Clinical Microbiology.

[7]  F. Tubau,et al.  Viridans Group Streptococci Are Donors in Horizontal Transfer of Topoisomerase IV Genes to Streptococcus pneumoniae , 2003, Antimicrobial Agents and Chemotherapy.

[8]  P. Huovinen,et al.  Antimicrobial Resistance of Invasive Pneumococci in Finland in 1999-2000 , 2003, Antimicrobial Agents and Chemotherapy.

[9]  R. Lütticken,et al.  Macrolide-Resistant Streptococcus pneumoniae and Streptococcus pyogenes in the Pediatric Population in Germany during 2000-2001 , 2003, Antimicrobial Agents and Chemotherapy.

[10]  M. P. Montanari,et al.  Phenotypes and Genotypes of Erythromycin-Resistant Pneumococci in Italy , 2003, Journal of Clinical Microbiology.

[11]  A. Pantosti,et al.  Inferring the potential success of pneumococcal vaccination in Italy: serotypes and antibiotic resistance of Streptococcus pneumoniae isolates from invasive diseases. , 2003, Microbial drug resistance.

[12]  M. Teuber,et al.  The Macrolide Efflux Genetic Assembly of Streptococcus pneumoniae Is Present in Erythromycin-Resistant Streptococcus salivarius , 2002, Antimicrobial Agents and Chemotherapy.

[13]  R. Leclercq,et al.  Resistance to Macrolides and Related Antibiotics in Streptococcus pneumoniae , 2002, Antimicrobial Agents and Chemotherapy.

[14]  G. Pozzi,et al.  Construction of new unencapsulated (rough) strains of Streptococcus pneumoniae. , 2002, Research in microbiology.

[15]  A. Pantosti,et al.  Macrolide Efflux Genes mef(A) and mef(E) Are Carried by Different Genetic Elements in Streptococcus pneumoniae , 2002, Journal of Clinical Microbiology.

[16]  G. Ippolito,et al.  Prevalence, Determinants, and Molecular Epidemiology of Streptococcus pneumoniae Isolates Colonizing the Nasopharynx of Healthy Children in Rome , 2002, European Journal of Clinical Microbiology and Infectious Diseases.

[17]  D. Stephens,et al.  Structure and dissemination of a chromosomal insertion element encoding macrolide efflux in Streptococcus pneumoniae. , 2001, The Journal of infectious diseases.

[18]  D. Hoban,et al.  Macrolide-Resistant Streptococcus pneumoniae in Canada during 1998–1999: Prevalence ofmef(A) and erm(B) and Susceptibilities to Ketolides , 2001, Antimicrobial Agents and Chemotherapy.

[19]  O. Cars,et al.  Variation in antibiotic use in the European Union , 2001, The Lancet.

[20]  A. Schuchat,et al.  The emergence of Streptococcus pneumoniae resistant to macrolide antimicrobial agents: a 6-year population-based assessment. , 2000, The Journal of infectious diseases.

[21]  G. Pozzi,et al.  Characterization of a Genetic Element Carrying the Macrolide Efflux Gene mef(A) in Streptococcus pneumoniae , 2000, Antimicrobial Agents and Chemotherapy.

[22]  P. Acar,et al.  Characterization of the Tn916-like Transposon Tn3872 in a Strain of Abiotrophia defectiva (Streptococcus defectivus) Causing Sequential Episodes of Endocarditis in a Child , 2000, Antimicrobial Agents and Chemotherapy.

[23]  K. Klugman,et al.  Acquisition of Chloramphenicol Resistance by the Linearization and Integration of the Entire Staphylococcal Plasmid pC194 into the Chromosome of Streptococcus pneumoniae , 2000, Antimicrobial Agents and Chemotherapy.

[24]  J. Rood,et al.  Nomenclature for Macrolide and Macrolide-Lincosamide-Streptogramin B Resistance Determinants , 1999, Antimicrobial Agents and Chemotherapy.

[25]  D. Mazel,et al.  Resistance gene capture. , 1999, Current opinion in microbiology.

[26]  M. Canron,et al.  Incidence of mefA and mefE Genes in Viridans Group Streptococci , 1999, Antimicrobial Agents and Chemotherapy.

[27]  Linda N. Lee,et al.  Detection of Tn917-Like Sequences within a Tn916-like Conjugative Transposon (Tn3872) in Erythromycin-Resistant Isolates of Streptococcus pneumoniae , 1998, Antimicrobial Agents and Chemotherapy.

[28]  L. Rice.  Tn916 Family Conjugative Transposons and Dissemination of Antimicrobial Resistance Determinants , 1998, Antimicrobial Agents and Chemotherapy.

[29]  J. Celli,et al.  Circularization of Tn916 is required for expression of the transposon‐encoded transfer functions: characterization of long tetracycline‐inducible transcripts reading through the attachment site , 1998, Molecular microbiology.

[30]  A. Kamath,et al.  Molecular cloning and functional analysis of a novel macrolide‐resistance determinant, mefA, from Streptococcus pyogenes , 1996, Molecular microbiology.

[31]  J. M. Smith,et al.  The tetracycline resistance gene tet(M) exhibits mosaic structure. , 1996, Plasmid.

[32]  G. Pozzi,et al.  Characterization of conjugative transposon Tn5251 of Streptococcus pneumoniae. , 1996, FEMS microbiology letters.

[33]  A. Stevens,et al.  Conjugative transposons: an unusual and diverse set of integrated gene transfer elements. , 1995, Microbiological reviews.

[34]  F. Bringel,et al.  Conjugative transposition of Tn916: preferred targets and evidence for conjugative transfer of a single strand and for a double‐stranded circular intermediate , 1994, Molecular microbiology.

[35]  Y. Su,et al.  Characterization of the left 4 kb of conjugative transposon Tn916: determinants involved in excision. , 1993, Plasmid.

[36]  C. Dowson,et al.  Evolution of penicillin resistance in Streptococcus pneumoniae; the role of Streptococcus mitis in the formation of a low affinity PBP2B in S. pneumoniae , 1993, Molecular microbiology.

[37]  D. Persing,et al.  Diagnostic molecular microbiology : principles and applications , 1993 .

[38]  P. Ayoubi,et al.  Tn5253, the pneumococcal omega (cat tet) BM6001 element, is a composite structure of two conjugative transposons, Tn5251 and Tn5252 , 1991, Journal of bacteriology.

[39]  W. R. Guild,et al.  Structure of a conjugative element in Streptococcus pneumoniae , 1986, Journal of bacteriology.

[40]  N. Shoemaker,et al.  Kinetics of integration of transforming DNA in pneumococcus. , 1972, Proceedings of the National Academy of Sciences of the United States of America.