Research Article: Identification of a Transcription Factor That Regulates Host Cell Exit and Virulence of Mycobacterium tuberculosis

Date Published: May 18, 2016

Publisher: Public Library of Science

Author(s): Lalitha Srinivasan, Serdar A. Gurses, Benjamin E. Hurley, Jessica L. Miller, Petros C. Karakousis, Volker Briken, Christopher M. Sassetti.

http://doi.org/10.1371/journal.ppat.1005652

Abstract

The interaction of Mycobacterium tuberculosis (Mtb) with host cell death signaling pathways is characterized by an initial anti-apoptotic phase followed by a pro-necrotic phase to allow for host cell exit of the bacteria. The bacterial modulators regulating necrosis induction are poorly understood. Here we describe the identification of a transcriptional repressor, Rv3167c responsible for regulating the escape of Mtb from the phagosome. Increased cytosolic localization of MtbΔRv3167c was accompanied by elevated levels of mitochondrial reactive oxygen species and reduced activation of the protein kinase Akt, and these events were critical for the induction of host cell necrosis and macroautophagy. The increase in necrosis led to an increase in bacterial virulence as reflected in higher bacterial burden and reduced survival of mice infected with MtbΔRv3167c. The regulon of Rv3167c thus contains the bacterial mediators involved in escape from the phagosome and host cell necrosis induction, both of which are crucial steps in the intracellular lifecycle and virulence of Mtb.

Partial Text

Apoptosis is a major programmed cell death pathway but now it is well established that necrosis can also be induced via defined signal transduction pathways [1,2]. The importance of apoptosis in host defense against pathogens is well described [3,4]. In contrast, the function of programmed necrosis in host resistance or susceptibility to pathogens is still an open question in many cases and may depend upon the context of the infection and the pathogen [5]. For instance, the RIPK1/3 necrosis pathway acts as a back-up mechanism of death induction in cells infected with viruses that are able to inhibit host cell apoptosis [6]. Consequently, programmed necrosis is associated with increased host resistance against viral pathogens in the case of vaccinia virus, adenovirus and MCMV [5,6]. Nevertheless, for the influenza A virus, programmed necrosis leads to increased pathology and host susceptibility [7]. Limited results are available for interaction of bacterial pathogens with host cell necrosis pathways but similar to viral pathogens the role of programmed necrosis may vary depending upon the pathogen. Enteropathogenic Escherichia coli can inhibit RIPK3-dependent necrosis via the glycosyl transferase NleB and this activity is important for bacterial virulence [8,9]. In contrast, IRF-3-dependent necrosis induction by Listeria monocytogenes promotes pathogen dissemination and virulence [10].

The intracellular location of bacteria has important consequences for their recognition by the host and the generation of innate and adaptive immune responses. Mtb was thought to restrict itself to a modified host cell phagosomal compartment after infection [79,80]. However, electron microscopy studies performed on infected macrophages and dendritic cells provided evidence that Mtb and other mycobacterial species are present in the cytosol and that phagosomal escape is dependent upon the ESX-1 secretion system [32,38]. This was confirmed by a FRET-based method dependent on β-lactamase production by Mtb in ex vivo-infected cells as well as in pulmonary phagocytic cells obtained from infected mice [33,34]. We report in the current study that the mycobacterial gene Rv3167c negatively regulates the escape of Mtb from the phagosome to the cytosol (Fig 4). Our study is the first to suggest that Mtb can exert temporal control on phagosomal escape as MtbΔRv3167c was found in the cytosol as early as 24h after infection while Mtb has been reported to access the cytosol much later in the infection process (4–5 days) [33]. We hypothesize that Rv3167c represses cytosolic escape at early stages when the bacterial load is low, favoring Mtb replication and establishment of infection. Genes involved in cytosolic escape could be induced once bacterial numbers reach about 20 per cell which seems to be an important threshold to switch on the cell escape program of Mtb [30]. Gene deletions resulting in increased cytosolic translocation from vacuolar compartments have been reported for other bacteria; for example, the sdhA (a Dot/Icm-secreted effector) mutant of Legionella pneumophila and the sifA (an SPI2-secreted effector) mutant of Salmonella typhimurium [81,82]. Deletion of the secreted phospholipase A abrogated the early escape of the L.pneumophila sdhA mutant [81]. The Mtb genome encodes four phospholipases (plc A-D), which could potentially contribute to the early escape of MtbΔRv3167c. The mechanisms involved in Mtb escape from the phagosome and eventual induction of host cell necrosis to exit the cell are difficult to study because of the slow kinetic of the process. Consequently, the early induction of cytosolic escape and necrosis by MtbΔRv3167c make it a useful model to study the host cell escape mechanisms of Mtb.

 

Source:

http://doi.org/10.1371/journal.ppat.1005652

 

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