Salmonella actively modulates TFEB in murine macrophages in a growth-phase and time-dependent manner

Abstract

ABSTRACT The transcription factor TFEB drives the expression of lysosomal, autophagic, and immune-responsive genes in response to LPS and phagocytosis. Interestingly, compounds that promote TFEB activity enhance bactericidal activity, while intracellular pathogens like Mycobacterium and Salmonella repress TFEB. However, Salmonella enterica sv. Typhimurium ( S . Typhimurium) was reported to actively stimulate TFEB, implying a benefit to Salmonella . To better understand the relationship between S . Typhimurium and TFEB, we assessed if S . Typhimurium regulated TFEB in macrophages in a manner dependent on infection conditions. We observed that macrophages that engulfed late-logarithmic grown Salmonella accumulated nuclear TFEB, comparable to macrophages that engulfed Escherichia coli . In contrast, stationary-phase S . Typhimurium infection of macrophages actively delayed TFEB nuclear mobilization. The delay in TFEB nuclear mobilization was not observed in macrophages that engulfed heat-killed stationary-phase Salmonella , or Salmonella lacking functional SPI-1 and SPI-2 type 3 secretion systems. S . Typhimurium mutated in the master virulence regulator phoP or the secreted effector genes sifA , and sopD also showed TFEB nuclear translocation. Interestingly, while E. coli survived better in tfeb −/− macrophages, S . Typhimurium growth was similar in wild-type and tfeb −/− macrophages. Moreover, Salmonella survival was not readily affected by its growth phase in wild-type or knockout macrophages, though in HeLa cells late-log Salmonella benefitted from the loss of TFEB. Priming macrophages with phagocytosis enhanced the killing of Salmonella in wild-type, but not in tfeb − /− macrophages. Collectively, S . Typhimurium orchestrate TFEB in a manner dependent on infection conditions, while disturbing this context-dependent control of TFEB may be detrimental to Salmonella survival. IMPORTANCE Activation of the host transcription factor TFEB helps mammalian cells adapt to stresses such as starvation and infection by upregulating lysosome, autophagy, and immuno-protective gene expression. Thus, TFEB is generally thought to protect host cells. However, it may also be that pathogenic bacteria like Salmonella orchestrate TFEB in a spatio-temporal manner to harness its functions to grow intracellularly. Indeed, the relationship between Salmonella and TFEB is controversial since some studies showed that Salmonella actively promotes TFEB, while others have observed that Salmonella degrades TFEB and that compounds that promote TFEB restrict bacterial growth. Our work provides a path to resolve these apparent discordant observations since we showed that stationary-grown Salmonella actively delays TFEB after infection, while late-log Salmonella is permissive of TFEB activation. Nevertheless, the exact function of this manipulation remains unclear, but conditions that erase the conditional control of TFEB by Salmonella may be detrimental to the microbe.