Mining of the Novel Virulent ATP-Binding Cassette Importers in Mycobacterium abscessus by Comparative Genomic Strategy

Abstract

Background:

The virulent ATP-binding cassette (ABC) importers from Mycobacterium abscessus, the most common native multidrug resistant and emerging opportunistic pathogen in rapidly growing NTM, were explored by comparative genomic study, in view of the fact that the ABC importers of Mycobacterium tuberculosis, responsible for uptaking metals, anions, amino acids, peptides, sugars, and other crucial substances from the host, had been proved to be closely related with the bacillus's virulence, survival in the host macrophages, antibiotic resistance, modulation of host immune system, and so on, although detailed mechanism was unclear.

Methods:

For virulent ABC importers from M. abscessus predicted by orthology and phylogeny analysis of nucleotide-binding domains (NBDs) of Mycobacterium smegmatis, M. abscessus, and M. tuberculosis, the antibiotic susceptibility of overexpression transformant and knockout mutant was assayed after confirmation by in vitro experiment.

Results:

Three-domain importers were dominant ones in M. abscessus (60.0%), four-domain ones dominant in M. tuberculosis (87.5%), whereas both types were same in M. smegmatis (41.9%). In the phylogenetic tree, the importers of M. abscessus (53.3%) and M. tuberculosis (62.5%) were mainly distributed in clay A, whereas the clay E was exclusively composed of M. smegmatis NBDs, which hinted possible reprogramming of the transporter system during the pathogen evolution. In clay A, MAB_2178 and others were predicted virulence-associated because of high sequence similarity to M. tuberculosis virulence importers.

Conclusions:

The importance and complexity of antibiotics resistance mechanisms of MAB_2176-2177-2178 were pointed out by its overexpression enhancing bacterial resistance to ciprofloxacin, clarithromycin, cefoxitin, and sensitivity to amikacin, and knockout having opposite phenotypes.