Polarly Localized EccE₁ Is Required for ESX-1 Function and Stabilization of ESX-1 Membrane Proteins in Mycobacterium...

Mycobacterium tuberculosis is a slow-growing intracellular bacterium with the ability to induce host cell death and persist indefinitely in the human body. This pathogen uses the specialized ESX-1 secretion system to secrete virulence factors and potent immunogenic effectors required for disease progression. ESX-1 is a multisubunit apparatus with a membrane complex that is predicted to form a channel in the cytoplasmic membrane. In M. tuberculosis this complex is composed of five membrane proteins: EccB₁, EccCa₁, EccCb₁, EccD₁, and EccE₁ In this study, we have characterized the membrane component EccE₁ and found that deletion of eccE₁ lowers the levels of EccB₁, EccCa₁, and EccD₁, thereby abolishing ESX-1 secretion and attenuating M. tuberculosisex vivo Surprisingly, secretion of EspB was not affected by loss of EccE₁ Furthermore, EccE₁ was found to be a membrane- and cell wall-associated protein that needs the presence of other ESX-1 components to assemble into a stable complex at the poles of M. tuberculosis Overall, this investigation provides new insights into the role of EccE₁ and its localization in M. tuberculosisIMPORTANCE Tuberculosis (TB), the world's leading cause of death of humans from an infectious disease, is caused by the intracellular bacterium Mycobacterium tuberculosis The development of successful strategies to control TB requires better understanding of the complex interactions between the pathogen and the human host. We investigated the contribution of EccE₁, a membrane protein, to the function of the ESX-1 secretion system, the major virulence determinant of M. tuberculosis By combining genetic analysis of selected mutants with eukaryotic cell biology and proteomics, we demonstrate that EccE₁ is critical for ESX-1 function, secretion of effector proteins, and pathogenesis. Our research improves knowledge of the molecular basis of M. tuberculosis virulence and enhances our understanding of pathogenesis.