Intrinsic rifamycin resistance of Mycobacterium abscessus is mediated by ADP-ribosyltransferase MAB_0591

Objectives: Rifampicin, a potent first-line TB drug of the rifamycin group, shows only little activity against the emerging pathogen Mycobacterium abscessus. Reportedly, bacterial resistance to rifampicin is associated with polymorphisms in the target gene rpoB or the presence of enzymes that modify and thereby inactivate rifampicin. The aim of this study was to investigate the role of the MAB_0591 (arr(Mab))-encoded rifampicin ADP-ribosyl-transferase (Arr_Mab) in innate high-level rifampicin resistance in M. abscessus. Methods: Recombinant Escherichia coli and Mycobacterium tuberculosis strains expressing MAB_0591 were generated, as was an M. abscessus deletion mutant deficient for MAB_0591. MIC assays were used to study susceptibility to rifampicin and C25 carbamate-modified rifamycin derivatives. Results: Heterologous expression of MAB_0591 conferred rifampicin resistance to E. coli and M. tuberculosis. Rifamycin MIC values were consistently lower for the M. abscessus Delta arr(Mab) mutant as compared with the M. abscessus ATCC 19977 parental type strain. The rifamycin WT phenotype was restored after complementation of the M. abscessus Delta arr(Mab) mutant with arr(Mab). Further MIC data demonstrated that a C25 modification increases rifamycin activity in WT M. abscessus. However, MIC studies in the M. abscessus Delta arr(Mab) mutant suggest that C25 modified rifamycins are still subject to modification by Arr_Mab. Conclusions: Our findings identify Arr_Mab as the major innate rifamycin resistance determinant of M. abscessus. Our data also indicate that Arr_Mab-mediated rifamycin resistance in M. abscessus can only in part be overcome by C25 carbamate modification.