Atypical Mycobacterium abscessus BlaRI Ortholog Mediates Regulation of Energy Metabolism but Not β‐Lactam Resistance

ABSTRACT Mycobacterium abscessus (Mab) is highly drug resistant, and understanding regulation of antibiotic resistance is critical to future antibiotic development. Regulatory mechanisms controlling Mab's β‐lactamase (BlaMab) that mediates β‐lactam resistance remain unknown. S. aureus encodes a prototypical protease‐mediated two‐component system BlaRI regulating the β‐lactamase BlaZ. BlaR binds extracellular β‐lactams, activating an intracellular peptidase domain which cleaves BlaI to derepress blaZ. Mycobacterium tuberculosis (Mtb) encodes homologs of BlaRI (which we will denote as BlaIR to reflect the inverted gene order in mycobacteria) that regulate not only the Mtb β‐lactamase, blaC, but also additional genes related to respiration. We identified orthologs of blaIRMtb in Mab and hypothesized that they regulate blaMab. Surprisingly, neither deletion of blaIRMab nor overexpression of only blaIMab altered blaMab expression or β‐lactam susceptibility. However, BlaIMab did bind to conserved motifs upstream of several Mab genes involved in respiration, yielding a putative regulon that partially overlapped with BlaIMtb. Prompted by evidence that respiration inhibitors including clofazimine induce the BlaI regulon in Mtb, we found that clofazimine triggers induction of blaIRMab and its downstream regulon. Highlighting an important role for BlaIRMab in adapting to disruptions in energy metabolism, constitutive repression of the BlaIMab regulon rendered Mab highly susceptible to clofazimine. In addition to our unexpected findings that BlaIRMab does not regulate β−lactam resistance, this study highlights the novel role of mycobacterial BlaRI‐type regulators in regulating electron transport and respiration. Mycobacterium abscessus (Mab) can cause skin infections and tuberculosis‐like lung disease. Mab has a reputation for extensive antibiotic resistance, so the poor understanding of regulation of antibiotic resistance and basic survival processes must be addressed. We explored the role of the regulatory proteins BlaIRMab, which had been shown to regulate antibiotic resistance in a species related to Mab, but unexpectedly found that BlaIRMab regulates many genes responsible for providing energy.