Mycobacterium tuberculosis FtsB and PerM interact via a C-terminal helix in FtsB to modulate cell division

Abstract

Latent infection by Mycobacterium tuberculosis (Mtb) impedes effective tuberculosis therapy and eradication. The protein PerM is essential for chronic Mtb infections in mice and acts via the divisome protein FtsB to modulate cell division. Using transgenic co-expression in Escherichia coli, we studied the Mtb PerM-FtsB interaction in isolation from other Mtb proteins, engineering PerM to enhance expression in the E. coli membrane. We confirmed the reported instability of Mtb FtsB, and we linked FtsB instability to a segment of FtsB predicted to bind cell-division proteins FtsL and FtsQ. Using fluorescence microscopy, we found that PerM stability hinged on its interaction with a C-terminal helix in FtsB. Molecular dynamics results supported the observation that FtsB stabilized PerM, and suggested that interactions at the PerM-FtsB interface differ from our initial structure prediction in a way that is consistent with PerM sequence conservation. Though narrowly conserved, the PerM-FtsB interaction emerges as a potential target for therapy targeting persistent infections by disrupting regulation of cell division. Integrating protein structure prediction, molecular dynamics and single-molecule microscopy, our approach is primed to screen potential inhibitors of the PerM-FtsB interaction and can be straightforwardly adapted to explore other putative interactions. ImportanceOur research reveals significant insights into the dynamic interaction between the proteins PerM and FtsB within Mycobacterium tuberculosis, contributing to our understanding of bacterial cell division mechanisms crucial for infection persistence. By combining innovative fluorescence microscopy and molecular dynamics, we established that the stability of these proteins is interdependent; molecular dynamics placing PerM-FtsB in the context of the mycobacterial divisome show how disrupting PerM-FtsB interaction can plausibly impact bacterial cell wall synthesis. These findings highlight the PerM-FtsB interface as a promising target for novel therapeutics aimed at combating persistent bacterial infections. Importantly, our approach can be adapted for similar studies in other bacterial systems, suggesting broad implications for microbial biology and antibiotic development.