TY - JOUR
T1 - PE-PGRS33 contributes to Mycobacterium tuberculosis entry in macrophages through interaction with TLR2
AU - Palucci, Ivana
AU - Sali, Michela
AU - Ria, Francesco
AU - Sanguinetti, Maurizio
AU - Delogu, Giovanni
AU - Camassa, Maria Serena
AU - Zumbo, Antonella
AU - Minerva, Mariachiara
AU - Iantomasi, Raffaella
AU - De Maio, Flavio
AU - Di Sante, Gabriele
AU - Cascioferro, Alessandro
AU - Anoosheh, Saber
AU - Palù, Giorgio
AU - Brennan, Michael J.
AU - Manganelli, Riccardo
PY - 2016
Y1 - 2016
N2 - PE-PGRS represent a large family of proteins typical of pathogenic mycobacteria whose members are characterized by an N-terminal PE domain followed by a large Gly-Ala repeat-rich C-terminal domain. Despite the abundance of PE-PGRS-coding genes in the Mycobacterium tuberculosis (Mtb) genome their role and function in the biology and pathogenesis still remains elusive. In this study, we generated and characterized an Mtb H37Rv mutant (MtbA33) in which the structural gene of PE-PGRS33, a prototypical member of the protein family, was inactivated. We showed that this mutant entered macrophages with an efficiency up to ten times lower than parental or complemented strains, while its efficiency in infecting pneumocytes remained unaffected. Interestingly, the lack of PE-PGRS33did not affect the intracellular growth of this mutant in macrophages. Using a series of functional deletion mutants of the PE-PGRS33 gene to complement the MtbA33 strain, we demonstrated that the PGRS domain is required to mediate cell entry into macrophages, with the key domain encompassing position 140-260 amino acids of PE-PGRS33. PE-PGRS33-mediated entry into macrophages was abolished in TLR2-deficient mice, as well as following treatment with wortmannin or an antibody against the complement receptor 3 (CR3), indicating that PE-PGRS33-mediated entry of Mtb in macrophages occurs through interaction with TLR2.
AB - PE-PGRS represent a large family of proteins typical of pathogenic mycobacteria whose members are characterized by an N-terminal PE domain followed by a large Gly-Ala repeat-rich C-terminal domain. Despite the abundance of PE-PGRS-coding genes in the Mycobacterium tuberculosis (Mtb) genome their role and function in the biology and pathogenesis still remains elusive. In this study, we generated and characterized an Mtb H37Rv mutant (MtbA33) in which the structural gene of PE-PGRS33, a prototypical member of the protein family, was inactivated. We showed that this mutant entered macrophages with an efficiency up to ten times lower than parental or complemented strains, while its efficiency in infecting pneumocytes remained unaffected. Interestingly, the lack of PE-PGRS33did not affect the intracellular growth of this mutant in macrophages. Using a series of functional deletion mutants of the PE-PGRS33 gene to complement the MtbA33 strain, we demonstrated that the PGRS domain is required to mediate cell entry into macrophages, with the key domain encompassing position 140-260 amino acids of PE-PGRS33. PE-PGRS33-mediated entry into macrophages was abolished in TLR2-deficient mice, as well as following treatment with wortmannin or an antibody against the complement receptor 3 (CR3), indicating that PE-PGRS33-mediated entry of Mtb in macrophages occurs through interaction with TLR2.
KW - Agricultural and Biological Sciences (all)
KW - Animals
KW - Bacterial Proteins
KW - Biochemistry, Genetics and Molecular Biology (all)
KW - Cell Line
KW - Macrophages
KW - Mice
KW - Mice, Inbred C57BL
KW - Mycobacterium tuberculosis
KW - Toll-Like Receptor 2
KW - Agricultural and Biological Sciences (all)
KW - Animals
KW - Bacterial Proteins
KW - Biochemistry, Genetics and Molecular Biology (all)
KW - Cell Line
KW - Macrophages
KW - Mice
KW - Mice, Inbred C57BL
KW - Mycobacterium tuberculosis
KW - Toll-Like Receptor 2
UR - http://hdl.handle.net/10807/117586
UR - http://www.plosone.org/article/fetchobject.action?uri=info:doi/10.1371/journal.pone.0150800&representation=pdf
U2 - 10.1371/journal.pone.0150800
DO - 10.1371/journal.pone.0150800
M3 - Article
VL - 11
SP - N/A-N/A
JO - PLoS One
JF - PLoS One
SN - 1932-6203
ER -