The Francis Crick Institute
anand-et-al-2023-er-dependent-membrane-repair-of-mycobacteria-induced-vacuole-damage.pdf (10.02 MB)

ER-dependent membrane repair of mycobacteria-induced vacuole damage.

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journal contribution
posted on 2023-11-02, 10:27 authored by Aby Anand, Anna-Carina Mazur, Patricia Rosell-Arevalo, Rico Franzkoch, Leonhard Breitsprecher, Stevanus A Listian, Sylvana V Hüttel, Danica Müller, Deise G Schäfer, Simone Vormittag, Hubert Hilbi, Markus Maniak, Maximiliano G Gutierrez, Caroline Barisch
Several intracellular pathogens, such as Mycobacterium tuberculosis, damage endomembranes to access the cytosol and subvert innate immune responses. The host counteracts endomembrane damage by recruiting repair machineries that retain the pathogen inside the vacuole. Here, we show that the endoplasmic reticulum (ER)-Golgi protein oxysterol-binding protein (OSBP) and its Dictyostelium discoideum homolog OSBP8 are recruited to the Mycobacterium-containing vacuole (MCV) dependent on the presence of the ESX-1 secretion system, suggesting that their mobilization is associated with membrane damage. Lack of OSBP8 causes a hyperaccumulation of phosphatidylinositol-4-phosphate (PI4P) on the MCV and decreased cell viability. OSBP8-depleted cells had reduced lysosomal and degradative capabilities of their vacuoles that favored mycobacterial growth. In agreement with a potential role of OSBP8 in membrane repair, human macrophages infected with M. tuberculosis recruited OSBP in an ESX-1-dependent manner. These findings identified an ER-dependent repair mechanism for restoring MCVs in which OSBP8 functions to equilibrate PI4P levels on damaged membranes. IMPORTANCE Tuberculosis still remains a global burden and is one of the top infectious diseases from a single pathogen. Mycobacterium tuberculosis, the causative agent, has perfected many ways to replicate and persist within its host. While mycobacteria induce vacuole damage to evade the toxic environment and eventually escape into the cytosol, the host recruits repair machineries to restore the MCV membrane. However, how lipids are delivered for membrane repair is poorly understood. Using advanced fluorescence imaging and volumetric correlative approaches, we demonstrate that this involves the recruitment of the endoplasmic reticulum (ER)-Golgi lipid transfer protein OSBP8 in the Dictyostelium discoideum/Mycobacterium marinum system. Strikingly, depletion of OSBP8 affects lysosomal function accelerating mycobacterial growth. This indicates that an ER-dependent repair pathway constitutes a host defense mechanism against intracellular pathogens such as M. tuberculosis.


Crick (Grant ID: CC2081, Grant title: Gutierrez CC2081) European Research Council (Grant ID: 772022 - DynaMO_TB, Grant title: ERC 772022 - DynaMO_TB)