Impact of protein domains on PE-PGRS30 polar localization in Mycobacteria

Flavio De Maio, Giuseppe Maulucci, Mariachiara Minerva, Saber Anoosheh, Ivana Palucci, Raffaella Iantomasi, Valentina Palmieri, Serena Camassa, Michela Sali, Maurizio Sanguinetti, Wilbert Bitter, Riccardo Manganelli, Marco De Spirito, Giovanni Delogu

Research output: Contribution to journalArticle

Abstract

PE-PGRS proteins are unique to the Mycobacterium tuberculosis complex and a number of other pathogenic mycobacteria. PE-PGRS30, which is required for the full virulence of M. tuberculosis (Mtb), has three main domains, i.e. an N-terminal PE domain, repetitive PGRS domain and the unique C-terminal domain. To investigate the role of these domains, we expressed a GFP-tagged PE-PGRS30 protein and a series of its functional deletion mutants in different mycobacterial species ( Mtb, Mycobacterium bovis BCG and Mycobacterium smegmatis ) and analysed protein localization by confocal microscopy. We show that PE-PGRS30 localizes at the mycobacterial cell poles in Mtb and M. bovis BCG but not in M. smegmatis and that the PGRS domain of the protein strongly contributes to protein cellular localization in Mtb. Immunofluorescence studies further showed that the unique C-terminal domain of PE-PGRS30 is not available on the surface, except when the PGRS domain is missing. Immunoblot demonstrated that the PGRS domain is required to maintain the protein strongly associated with the non-soluble cellular fraction. These results suggest that the repetitive GGA-GGN repeats of the PGRS domain contain specific sequences that contribute to protein cellular localization and that polar localization might be a key step in the PE-PGRS30-dependent virulence mechanism.

Original languageEnglish
Article numbere112482
JournalPLoS One
Volume9
Issue number11
DOIs
Publication statusPublished - Nov 12 2014

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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    De Maio, F., Maulucci, G., Minerva, M., Anoosheh, S., Palucci, I., Iantomasi, R., Palmieri, V., Camassa, S., Sali, M., Sanguinetti, M., Bitter, W., Manganelli, R., De Spirito, M., & Delogu, G. (2014). Impact of protein domains on PE-PGRS30 polar localization in Mycobacteria. PLoS One, 9(11), [e112482]. https://doi.org/10.1371/journal.pone.0112482