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bioRxiv · 10.1101/2023.02.13.528419

The SapM phosphatase arrests phagosome maturation in an ESX-1 independent manner in Mycobacterium tuberculosis and BCG

Xander, C.·Rajagopalan, S.·Jacobs, W. R.·Braunstein, M.
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Abstract

Mycobacterium tuberculosis (Mtb) is an intracellular pathogen that survives and grows in macrophages. A mechanism used by Mtb to achieve intracellular survival is to secrete effector molecules that arrest the normal process of phagosome maturation. Through phagosome maturation arrest (PMA), Mtb remains in an early phagosome and avoids delivery to degradative phagolysosomes. One PMA effector of Mtb is the secreted SapM phosphatase. Because the host target of SapM, phosphatidylinositol-3-phosphate (PI3P), is located on the cytosolic face of the phagosome, SapM needs to be both released by the mycobacteria and escape the phagosome to carry out its function. To date, the only mechanism known for Mtb molecules to escape the phagosome is phagosome permeabilization by the ESX-1 secretion system. To understand this step of SapM function in PMA, we generated identical in-frame sapM mutants in both the attenuated Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccine strain, which lacks the ESX-1 system, and Mtb. Characterization of these mutants demonstrated that SapM is required for PMA in both BCG and Mtb. Further, by establishing a role for SapM in PMA in BCG, and subsequently in a Mtb mutant lacking the ESX-1 system, we demonstrated that the role of SapM is ESX-1-independent. We further determined that ESX-2 or ESX-4 are also not required for SapM to function in PMA. These results indicate that SapM is a secreted effector of PMA in both BCG and Mtb and that it functions independent of the known mechanism for Mtb molecules to escape the phagosome.

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Xander, C., Rajagopalan, S., Jacobs, W. R., Braunstein, M.. 2023-02-14. The SapM phosphatase arrests phagosome maturation in an ESX-1 independent manner in Mycobacterium tuberculosis and BCG. https://doi.org/10.1101/2023.02.13.528419

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