Identification of a novel tetracycline resistance gene, tet(63), located on a multiresistance plasmid from Staphylococcus aureus

2020 ◽  
Author(s):  
Yao Zhu ◽  
Changzhen Wang ◽  
Stefan Schwarz ◽  
Wenyu Liu ◽  
Qin Yang ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Resistance Gene ◽  
Efflux Pump ◽  
Tetracycline Resistance ◽  
Major Facilitator Superfamily ◽  
Whole Genome Sequence ◽  
Tetracycline Resistance Gene ◽  
Efflux Pump Inhibitor ◽  
Illumina Hiseq

Abstract Objectives To identify and characterize a novel tetracycline resistance gene on a multiresistance plasmid from Staphylococcus aureus SA01 of chicken origin. Methods MICs were determined by broth microdilution according to CLSI recommendations. The whole genome sequence of S. aureus SA01 was determined via Illumina HiSeq and Oxford Nanopore platforms followed by a hybrid assembly. The new tet gene was cloned and expressed in S. aureus. The functionality of the corresponding protein as an efflux pump was tested by efflux pump inhibition assays. Results A novel tetracycline resistance gene, tet(63), was identified on a plasmid in S. aureus SA01. The cloned tet(63) gene was functionally expressed in S. aureus and shown to confer resistance to tetracycline and doxycycline, and a slightly elevated MIC of minocycline. The tet(63) gene encodes a 459 amino acid efflux protein of the major facilitator superfamily that consists of 14 predicted transmembrane helices. The results of efflux pump inhibitor assays confirmed the function of Tet(63) as an efflux protein. The deduced amino acid sequence of the Tet(63) protein exhibited 73.0% identity to the tetracycline efflux protein Tet(K). The plasmid pSA01-tet, on which tet(63) was located, had a size of 25664 bp and also carried the resistance genes aadD, aacA-aphD and erm(C). Conclusions A novel tetracycline resistance gene, tet(63), was identified in S. aureus. Its location on a multiresistance plasmid might support the co-selection of tet(63) under the selective pressure imposed by the use of macrolides, lincosamides and aminoglycosides.

scholarly journals Identification of a Novel Trimethoprim Resistance Gene, dfrK, in a Methicillin-Resistant Staphylococcus aureus ST398 Strain and Its Physical Linkage to the Tetracycline Resistance Gene tet(L)

2008 ◽  
Vol 53 (2) ◽  
pp. 776-778 ◽  
Author(s):  
Kristina Kadlec ◽  
Stefan Schwarz
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Gene ◽  
Dihydrofolate Reductase ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Tetracycline Resistance ◽  
Sequence Type ◽  
Tetracycline Resistance Gene ◽  
Close Proximity ◽  
Physical Linkage ◽  
Dihydrofolate Reductases

ABSTRACT A novel trimethoprim resistance gene, designated dfrK, was detected in close proximity to the tetracycline resistance gene tet(L) on the ca. 40-kb plasmid pKKS2187 in a porcine methicillin (meticillin)-resistant Staphylococcus aureus isolate of sequence type 398. The dfrK gene encodes a 163-amino-acid dihydrofolate reductase that differs from all so-far-known dihydrofolate reductases.

scholarly journals An IS257-Derived Hybrid Promoter Directs Transcription of a tetA(K) Tetracycline Resistance Gene in theStaphylococcus aureus Chromosomal mecRegion

2000 ◽  
Vol 182 (12) ◽  
pp. 3345-3352 ◽  
Author(s):  
Alice E. Simpson ◽  
Ronald A. Skurray ◽  
Neville Firth
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Gene ◽  
Gene Dosage ◽  
Tetracycline Resistance ◽  
Resistance Determinant ◽  
Tetracycline Resistance Gene ◽  
Methicillin Resistant ◽  
Hybrid Promoter ◽  
Genetic Rearrangements

ABSTRACT Transcription of the tetA(K) tetracycline resistance determinant encoded by an IS257-flanked cointegrated copy of a pT181-like plasmid, located within the chromosomal mecregion of a methicillin-resistant Staphylococcus aureusisolate, has been investigated. The results demonstrated that transcription of tetA(K) in this strain is directed by both an IS257-derived hybrid promoter, which is stronger than the native tetA(K) promoter in the autonomous form of pT181, and a complete outwardly directed promoter identified within one end of IS257. Despite lower gene dosage, the chromosomal configuration was shown to afford a higher level of resistance than that mediated by pT181 in an autonomous multicopy state. Furthermore, competition studies revealed that a strain carrying the chromosomaltetA(K) determinant exhibited a higher level of fitness in the presence of tetracycline but not in its absence. This finding suggests that tetracycline has been a selective factor in the emergence of strains carrying a cointegrated pT181-like plasmid in their chromosomes. The results highlight the potential of IS257to influence the expression of neighboring genes, a property likely to enhance its capacity to mediate advantageous genetic rearrangements.

scholarly journals Identification of a New Ribosomal Protection Type of Tetracycline Resistance Gene, tet(36), from Swine Manure Pits

2003 ◽  
Vol 69 (7) ◽  
pp. 4151-4158 ◽  
Author(s):  
Gabrielle Whittle ◽  
Terence R. Whitehead ◽  
Nathan Hamburger ◽  
Nadja B. Shoemaker ◽  
Michael A. Cotta ◽  
...  
Keyword(s):  
Amino Acid ◽  
Resistance Gene ◽  
Anaerobic Bacteria ◽  
Swine Manure ◽  
Tetracycline Resistance ◽  
Amino Acid Identity ◽  
Phylogenetic Groups ◽  
Tetracycline Resistance Gene ◽  
Acid Identity ◽  
Swine Feces

ABSTRACT Previously, only one ribosome protection type of a tetracycline resistance gene, tetQ, had been identified in Bacteroides spp. During an investigation of anaerobic bacteria present in swine feces and manure storage pits, a tetracycline-resistant Bacteroides strain was isolated. Subsequent analysis showed that this new Bacteroides strain, Bacteroides sp. strain 139, did not contain tetQ but contained a previously unidentified tetracycline resistance gene. Sequence analysis showed that the tetracycline resistance gene from Bacteroides sp. strain 139 encoded a protein (designated Tet 36) that defines a new class of ribosome protection types of tetracycline resistance. Tet 36 has 60% amino acid identity over 640 aa to TetQ and between 31 and 49% amino acid identity to the nine other ribosome protection types of tetracycline resistance genes. The tet(36) region was not observed to transfer from Bacteroides sp. strain 139 to another Bacteroides sp. under laboratory conditions. Yet tet(36) was found in other genera of bacteria isolated from the same swine manure pits and from swine feces. Phylogenetic analysis of the tet(36)-containing isolates indicated that tet(36) was present not only in the Cytophaga-Flavobacter-Bacteroides group to which Bacteroides sp. strain 139 belongs but also in gram-positive genera and gram-negative proteobacteria, indicating that horizontal transfer of tet(36) is occurring between these divergent phylogenetic groups in the farm environment.

scholarly journals Copresence oftet(K) andtet(M) in Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Is Associated with Increased Fitness during Exposure to Sublethal Concentrations of Tetracycline

2016 ◽  
Vol 60 (7) ◽  
pp. 4401-4403 ◽  
Author(s):  
Jesper Larsen ◽  
Julie Clasen ◽  
Julie E. Hansen ◽  
Wilhelm Paulander ◽  
Andreas Petersen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Gene ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Clonal Complex ◽  
Tetracycline Resistance ◽  
Tetracycline Resistance Gene ◽  
Methicillin Resistant ◽  
Content Type ◽  
Sublethal Concentrations ◽  
Staphylococcal Cassette Chromosome

ABSTRACTThe tetracycline resistance genetet(K) was shown to be integrated within the predominant staphylococcal cassette chromosomemec(SCCmec) element of Danish livestock-associated methicillin-resistantStaphylococcus aureusCC398 (LA-MRSA CC398). These LA-MRSA CC398 isolates already possessedtet(M), but the acquisition oftet(K) significantly improved their fitness at sublethal concentrations of tetracycline. Becausetet(K) is genetically linked to SCCmec, the use of tetracycline in food animals may have contributed to the successful spread of LA-MRSA CC398.

scholarly journals The TetA(K) Tetracycline/H+ Antiporter from Staphylococcus aureus: Mutagenesis and Functional Analysis of Motif C

2000 ◽  
Vol 182 (6) ◽  
pp. 1492-1498 ◽  
Author(s):  
Samantha L. Ginn ◽  
Melissa H. Brown ◽  
Ronald A. Skurray
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Tetracycline Resistance ◽  
Transport Proteins ◽  
Major Facilitator Superfamily ◽  
Site Directed Mutagenesis ◽  
Transport Activity ◽  
Transmembrane Segment ◽  
Mutant Proteins ◽  
Major Facilitator

ABSTRACT Conserved motif C, identified within members of the major facilitator superfamily (MFS) of transport proteins that mediate drug export, was examined in the tetracycline resistance efflux protein TetA(K) from Staphylococcus aureus; motif C is contained within transmembrane segment 5. Using site-directed mutagenesis, the importance of the conserved glycine (G151, G155, G159, and G160) and proline (P156) residues within this motif was investigated. Over 40 individual amino acid replacements were introduced; however, only alanine and serine substitutions for glycine at G151, G155, and G160 were found to retain significant levels of tetracycline resistance and transport activity in cells expressing mutant proteins. Notably, P156 and G159 appear to be crucial, as amino acid replacements at these positions either significantly reduced or abolished tetracycline/H+ activity. The highly conserved nature of motif C and its distribution throughout drug exporters imply that the residues of motif C play a similar role in all MFS proteins that function as antiporters.

scholarly journals Staphylococcus aureus Mutants Isolated via Exposure to Nonfluorinated Quinolones: Detection of Known and Unique Mutations

2001 ◽  
Vol 45 (12) ◽  
pp. 3422-3426 ◽  
Author(s):  
Siddhartha Roychoudhury ◽  
Tracy L. Twinem ◽  
Kelly M. Makin ◽  
Mark A. Nienaber ◽  
Chuiying Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Sequence Analysis ◽  
Efflux Pump ◽  
Serial Passage ◽  
Efflux Pump Inhibitor ◽  
In Vitro Development ◽  
Development Of Resistance ◽  
High Level ◽  
Vitro Development

ABSTRACT The in vitro development of resistance to the new nonfluorinated quinolones (NFQs; PGE 9262932, PGE 4175997, and PGE 9509924) was investigated in Staphylococcus aureus. At concentrations two times the MIC, step 1 mutants were isolated more frequently with ciprofloxacin and trovafloxacin (9.1 × 10−8 and 5.7 × 10−9, respectively) than with the NFQs, gatifloxacin, or clinafloxacin (<5.7 × 10−10). Step 2 and step 3 mutants were selected via exposure of a step 1 mutant (selected with trovafloxacin) to four times the MICs of trovafloxacin and PGE 9262932. The step 1 mutant contained the known Ser80-Phe mutation in GrlA, and the step 2 and step 3 mutants contained the known Ser80-Phe and Ser84-Leu mutations in GrlA and GyrA, respectively. Compared to ciprofloxacin, the NFQs were 8-fold more potent against the parent and 16- to 128-fold more potent against the step 3 mutants. Mutants with high-level NFQ resistance (MIC, 32 μg/ml) were isolated by the spiral plater-based serial passage technique. DNA sequence analysis of three such mutants revealed the following mutations: (i) Ser84-Leu in GyrA and Glu84-Lys and His103-Tyr in GrlA; (ii) Ser-84Leu in GyrA, Ser52-Arg in GrlA, and Glu472-Val in GrlB; and (iii) Ser84-Leu in GyrA, Glu477-Val in GyrB, and Glu84-Lys and His103-Tyr in GrlA. Addition of the efflux pump inhibitor reserpine (10 μg/ml) resulted in 4- to 16-fold increases in the potencies of the NFQs against these mutants, whereas it resulted in 2-fold increases in the potencies of the NFQs against the parent.

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