Mutations outside the rifampicin resistance-determining region associated with rifampicin resistance in Mycobacterium tuberculosis

Kit Hang Siu, Ying Zhang, Terrence C K Lau, Ricky W T Lau, Pak Leung Ho, Wing Wai Yew, Stephen K W Tsui, Vincent C C Cheng, Kwok Yung Yuen, Wing Cheong Yam

Research output: Journal article publicationJournal articleAcademic researchpeer-review

63 Citations (Scopus)

Abstract

Objectives: Ninety-six percent of rifampicin resistance in Mycobacterium tuberculosis was shown to be associated with mutations inside the 81 bp rifampicin resistance-determining region (RRDR) located in the centre of the rpoB gene. The detection of rifampicin resistance by targeting the RRDR failed to match with a resistant phenotype in 4% of all cases. Our study aims to identify the mutations outside the RRDR that are associated with rifampicin resistance in M. tuberculosis. Methods and results: Among 50 rifampicin-resistant and 20 rifampicin-susceptible clinical isolates of M. tuberculosis, 2 of the rifampicin-resistant isolates did not harbour any known mutations in the RRDR. Sequencing analysis of the whole rpoB gene identified two rare mutations, V146F and I572F. A molecular structure model based on Thermus thermophilus RpoB revealed that both these substituted amino acids are located in close proximity to the rifampicin-binding pocket of the β-subunit. Substitutions of simple amino acids for bulky ones are likely to affect the protein-drug interaction. Cloning and transformation of the mutated rpoB gene into wild-type Mycobacterium smegmatis and M. tuberculosis successfully elevated the MIC of rifampicin and conferred the rifampicin resistance phenotype. Conclusions: Our study showed that amino acid positions 146 and 572 are associated with rifampicin resistance in M. tuberculosis in addition to the RRDR. Molecular assays for identifying rifampicin-resistant M. tuberculosis might be improved in terms of accuracy by including these two positions.
Original languageEnglish
Article numberdkq519
Pages (from-to)730-733
Number of pages4
JournalJournal of Antimicrobial Chemotherapy
Volume66
Issue number4
DOIs
Publication statusPublished - 1 Apr 2011
Externally publishedYes

Keywords

  • Mycobacterial RNA polymerase
  • Mycobacterium smegmatis
  • RpoB transformation

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

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