title: Mycobacterium tuberculosis EsxO (Rv2346c) promotes bacillary survival by inducing oxidative stress mediated genomic instability in macrophages.
abstract: Mycobacterium tuberculosis (Mtb) survives inside the macrophages by modulating the host immune responses in its favor. The 6-kDa early secretory antigenic target (ESAT-6; esxA) of Mtb is known as a potent virulence and T-cell antigenic determinant. At least 23 such ESAT-6 family proteins are encoded in the genome of Mtb; however, the function of many of them is still unknown. We herein report that ectopic expression of Mtb Rv2346c (esxO), a member of ESAT-6 family proteins, in non-pathogenic Mycobacterium smegmatis strain (MsmRv2346c) aids host cell invasion and intracellular bacillary persistence. Further mechanistic studies revealed that MsmRv2346c infection abated macrophage immunity by inducing host cell death and genomic instability as evident from the appearance of several DNA damage markers. We further report that the induction of genomic instability in infected cells was due to increase in the hosts oxidative stress responses. MsmRv2346c infection was also found to induce autophagy and modulate the immune function of macrophages. In contrast, blockade of Rv2346c induced oxidative stress by treatment with ROS inhibitor N-acetyl-L-cysteine prevented the host cell death, autophagy induction and genomic instability in infected macrophages. Conversely, MtbΔRv2346c mutant did not show any difference in intracellular survival and oxidative stress responses. We envision that Mtb ESAT-6 family protein Rv2346c dampens antibacterial effector functions namely by inducing oxidative stress mediated genomic instability in infected macrophages, while loss of Rv2346c gene function may be compensated by other redundant ESAT-6 family proteins. Thus EsxO plays an important role in mycobacterial pathogenesis in the context of innate immunity.
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
authors.forename: Soumitra
authors.initials: S
authors.lastname: Mohanty
authors.affiliation: Institute of Medical Microbiology, University of Zurich, Gloriastrasse 32, CH 8006 Zurich, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Dal Molin
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
authors.forename: Geetanjali
authors.initials: G
authors.lastname: Ganguli
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
authors.forename: Avinash
authors.initials: A
authors.lastname: Padhi
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
authors.forename: Prajna
authors.initials: P
authors.lastname: Jena
authors.affiliation: Institute of Medical Microbiology, University of Zurich, Gloriastrasse 32, CH 8006 Zurich, Switzerland.
authors.forename: Petra
authors.initials: P
authors.lastname: Selchow
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
authors.forename: Srabasti
authors.initials: S
authors.lastname: Sengupta
authors.affiliation: Institute of Medical Microbiology, University of Zurich, Gloriastrasse 32, CH 8006 Zurich, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Meuli
authors.affiliation: Institute of Medical Microbiology, University of Zurich, Gloriastrasse 32, CH 8006 Zurich, Switzerland; National Reference Laboratory for Mycobacteria, Gloriastrasse 30, CH 8006 Zurich, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Sander
authors.affiliation: School of Biotechnology, KIIT University, Bhubaneswar, Odisha, India. Electronic address: asonawane@kiitbiotech.ac.in.
authors.forename: Avinash
authors.initials: A
authors.lastname: Sonawane
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001343:Autophagy; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004249:DNA Damage; D005260:Female; D042822:Genomic Instability; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D007113:Immunity, Innate; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D050296:Microbial Viability; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000472:pathogenicity; D018384:Oxidative Stress / Q000187:drug effects; D000067996:RAW 264.7 Cells; D013997:Time Factors; D014774:Virulence
chemical_list: D000975:Antioxidants; D001426:Bacterial Proteins; D019883:Alkyl and Aryl Transferases; C499606:Rv2361c protein, Mycobacterium tuberculosis
keywords: Genomic instability; Macrophages; Mycobacterium tuberculosis; Oxidative stress; Rv2346c; Survival; esxO