title: Disruption of the ESX-5 system of Mycobacterium tuberculosis causes loss of PPE protein secretion, reduction of cell wall integrity and strong attenuation.
abstract: The chromosome of Mycobacterium tuberculosis encodes five type VII secretion systems (ESX-1-ESX-5). While the role of the ESX-1 and ESX-3 systems in M. tuberculosis has been elucidated, predictions for the function of the ESX-5 system came from data obtained in Mycobacterium marinum, where it transports PPE and PE_PGRS proteins and modulates innate immune responses. To define the role of the ESX-5 system in M. tuberculosis, in this study, we have constructed five M. tuberculosis H37Rv ESX-5 knockout/deletion mutants, inactivating eccA(5), eccD(5), rv1794 and esxM genes or the ppe25-pe19 region. Whereas the Mtbrv1794ko displayed no obvious phenotype, the other four mutants showed defects in secretion of the ESX-5-encoded EsxN and PPE41, a representative member of the large PPE protein family. Strikingly, the MtbeccD(5) ko mutant also showed enhanced sensitivity to detergents and hydrophilic antibiotics. When the virulence of the five mutants was evaluated, the MtbeccD(5) ko and MtbΔppe25-pe19 mutants were found attenuated both in macrophages and in the severe combined immune-deficient mouse infection model. Altogether these findings indicate an essential role of ESX-5 for transport of PPE proteins, cell wall integrity and full virulence of M. tuberculosis, thereby opening interesting new perspectives for the study of this human pathogen.
authors.affiliation: Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, University of Pisa, Pisa, Italy. d.bottai@biomed.unipi.it
authors.forename: Daria
authors.initials: D
authors.lastname: Bottai
authors.forename: Mariagrazia
authors.initials: M
authors.lastname: Di Luca
authors.forename: Laleh
authors.initials: L
authors.lastname: Majlessi
authors.forename: Wafa
authors.initials: W
authors.lastname: Frigui
authors.forename: Roxane
authors.initials: R
authors.lastname: Simeone
authors.forename: Fadel
authors.initials: F
authors.lastname: Sayes
authors.forename: Wilbert
authors.initials: W
authors.lastname: Bitter
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Brennan
authors.forename: Claude
authors.initials: C
authors.lastname: Leclerc
authors.forename: Giovanna
authors.initials: G
authors.lastname: Batoni
authors.forename: Mario
authors.initials: M
authors.lastname: Campa
authors.forename: Roland
authors.initials: R
authors.lastname: Brosch
authors.forename: Semih
authors.initials: S
authors.lastname: Esin
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D002473:Cell Wall / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D016513:Mice, SCID; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D021381:Protein Transport; D014376:Tuberculosis / Q000382:microbiology*; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems