Molecular basis of streptomycin resistance in <i>Mycobacterium tuberculosis</i>: alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosomal RNA pseudoknot

Finken, Marion; Kirschner, Philip; Meier, A; Wrede, Annette; Böttger, Erik C.

doi:10.1111/j.1365-2958.1993.tb01253.x

Cited by 328 publications

(225 citation statements)

References 34 publications

Supporting

Mentioning

209

Contrasting

Unclassified

Order By: Relevance

“…Thus, the loops of 16S rRNA that interact with the S12 protein constitute an easily selected mutation site. Such rrs mutations are clustered in the highly conserved 530 loop and in the adjacent 915 region (6). In addition, a 1400A3G mutation of rrs was identified in both amikacinand kanamycin-resistant clinical isolates of M. tuberculosis (1,29).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Detection of Multidrug Resistance in Mycobacterium tuberculosis

et al. 2007

View full text Add to dashboard Cite

We developed a DNA sequencing-based method to detect mutations in the genome of drug-resistant Mycobacterium tuberculosis. Drug resistance in M. tuberculosis is caused by mutations in restricted regions of the genome. Eight genome regions associated with drug resistance, including rpoB for rifampin (RIF), katG and the mabA (fabG1)-inhA promoter for isoniazid (INH), embB for ethambutol (EMB), pncA for pyrazinamide (PZA), rpsL and rrs for streptomycin (STR), and gyrA for levofloxacin, were amplified simultaneously by PCR, and the DNA sequences were determined. It took 6.5 h to complete all procedures. Among the 138 clinical isolates tested, 55 were resistant to at least one drug. Thirty-four of 38 INH-resistant isolates (89.5%), 28 of 28 RIF-resistant isolates (100%), 15 of 18 EMB-resistant isolates (83.3%), 18 of 30 STR-resistant isolates (60%), and 17 of 17 PZA-resistant isolates (100%) had mutations related to specific drug resistance. Eighteen of these mutations had not been reported previously. These novel mutations include one in rpoB, eight in katG, one in the mabA-inhA regulatory region, two in embB, five in pncA, and one in rrs. Escherichia coli isolates expressing individually five of the eight katG mutations showed loss of catalase and INH oxidation activities, and isolates carrying any of the five pncA mutations showed no pyrazinamidase activity, indicating that these mutations are associated with INH and PZA resistance, respectively. Our sequencing-based method was also useful for testing sputa from tuberculosis patients and for screening of mutations in Mycobacterium bovis. In conclusion, our new method is useful for rapid detection of multiple-drug-resistant M. tuberculosis and for identifying novel mutations in drug-resistant M. tuberculosis.The emergence and spread of drug-resistant strains of Mycobacterium tuberculosis, especially multidrug-resistant (MDR) strains, are serious threats to the control of tuberculosis and comprise an increasing public health problem (40). Patients infected with MDR strains, which are defined as strains resistant to both rifampin (RIF) and isoniazid (INH), are difficult to cure and are more likely to remain sources of infection for a longer period of time than are patients with drug-susceptible strains (40).It is essential that rapid drug susceptibility tests be developed to prevent the spread of MDR M. tuberculosis. The time necessary for culture of specimens was reduced by the radiometric BACTEC 460TB system (BD Biosciences, Sparks, MD), the nonradiometric ESP II system (Trek Diagnostics, Westlake, OH), and other rapid broth methods, such as BACTEC MGIT 960 SIRE (BD Biosciences) (20). These drug susceptibility tests, however, still require 1 to 2 weeks for final determination and reporting to the clinician (23). Additional reductions in the detection period are needed.Drug resistance in M. tuberculosis is caused by mutations in relatively restricted regions of the genome (17, 39). Mutations associated with drug resistance occur in rpoB for RIF, katG and the promote...

show abstract

Section: Discussionmentioning

confidence: 99%

“…M. tuberculosis becomes resistant when targets of STR in the ribosomes are mutated. The principal site of mutation is the rpsL gene, which encodes ribosomal protein S12 (6,19,27). The most frequently observed mutation in rpsL was K43R.…”

Section: Discussionmentioning

confidence: 99%

Detection of Multidrug Resistance in Mycobacterium tuberculosis

et al. 2007

View full text Add to dashboard Cite

show abstract

“…32 Essas mutações freqüentemente estão associadas à perda da atividade de catalaseperoxidase. 9,11,27,40 Essa enzima é responsável também pela ativação da INH endogenamente.…”

Section: Fármacosunclassified

“…12 A resistência à SM em M. tuberculosis ocorre por mutações no alvo do fármaco, mais especificamente nos ribossomos. O principal sítio de mutação é o gene rpsL, que codifica a proteína ribossomal S12, 5,11,15,33 em que ocorrem mutações resultando na substituição de um único aminoácido. O mapeamento dessas mutações revela que estas ocorrem em regiões altamente conservadas do gene.…”

Section: Resistência à Estreptomicinaunclassified

Tuberculose resistente: revisão molecular

et al. 2002

View full text Add to dashboard Cite

ResumoO progresso na compreensão dos mecanismos de resistência aos fármacos usados no tratamento da tuberculose tem permitido o desenvolvimento de novos métodos para a detecção da tuberculose resistente. A resistente aos fármacos representa uma ameaça para os programas de controle da tuberculose. Para tanto, é necessário conhecer o padrão de sensibilidade das linhagens para fornecer o tratamento adequado. Os estudos moleculares dos mecanismos de ação dos fármacos antituberculose têm elucidado as bases genéticas da resistência aos fármacos em Mycobacterium tuberculosis. Os mecanismos de resistência aos fármacos na tuberculose são causados por mutações cromossomais em diferentes genes da bactéria. Durante a exposição aos fármacos, há uma pressão seletiva favorecendo o desenvolvimento de linhagens resistentes. A tuberculose multirresistente é um problema nacional e internacional que traz sérias dificuldades para o controle global da doença. Realizou-se uma revisão sobre os mecanismos moleculares associados à resistência aos fármacos com ênfase nas novas perspectivas para detectar os isolados resistentes. AbstractProgress to understanding the basis of resistance to antituberculous drugs has allowed molecular tests for detection of drug-resistant tuberculosis to be developed. Drug-resistant tuberculosis poses a threat to tuberculosis control programs. It is necessary thus to know drug susceptibilities of individual patient's strain to provide the appropriate drug combinations. Molecular studies on the mechanism of action of antituberculous drugs have elucidated the genetic basis of drug resistance in M. tuberculosis. The mechanisms of drug resistance in tuberculosis are a result of chromosomal mutations in different genes of the bacteria. Upon drug exposure there is a selective pressure for such resistant mutants. Multidrug-resistant tuberculosis is a health problem of increasing significance for the whole global community. This paper reviews the molecular mechanisms associated with drugresistance as well the new perspectives for detecting resistant isolates. Current Comments Atualização

show abstract

“…The construction of the M. bovis BCG library has been described elsewhere (Finken et al, 1993). A genomic library of M. smegmatis was constructed in a similar manner.…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

The role of ribosomal RNAs in macrolide resistance

Sander

Prammananan

Meier

et al. 1997

Molecular Microbiology

Self Cite

View full text Add to dashboard Cite

SummaryMacrolides are bacteriostatic antibiotics which interfere with the peptidyltransfer function of the ribosome. We have investigated the molecular mechanisms underlying macrolide resistance in Mycobacterium smegmatis, an eubacterium carrying two rRNA operons. Surprisingly, drug resistance was associated not with alterations in ribosomal proteins, but with a single point mutation in the peptidyltransferase region of one of the two 23S RNA genes, i.e. A2058→G or A2059→G. This mutation resulted in a heterozygous organism with a mutated and a wild-type rRNA operon respectively. Reverse transcriptase sequencing indicated the expression of both wild-type and mutated rRNAs. The mutated operon was introduced into genetically engineered rrn ¹ strains of M. smegmatis carrying a single functional rRNA operon and into parental M. smegmatis with two chromosomal rRNA operons, using gene transfer as well as gene replacement techniques. The results obtained demonstrate the dominant nature of resistance. As exemplified in our results on macrolide resistance, a complete set of genetic tools is now available, which allows questions of dominance vs. recessivity and gene dosage effects in eubacterial ribosomal nucleic acids to be addressed experimentally in vivo.

show abstract

Molecular basis of streptomycin resistance in Mycobacterium tuberculosis: alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosomal RNA pseudoknot

Cited by 328 publications

References 34 publications

Detection of Multidrug Resistance in Mycobacterium tuberculosis

Detection of Multidrug Resistance in Mycobacterium tuberculosis

Tuberculose resistente: revisão molecular

The role of ribosomal RNAs in macrolide resistance

Contact Info

Product

Resources

About