Functional dissection of the pe domain responsible for translocation of PE_PGRS33 across the mycobacterial cell wall

Alessandro Cascioferro, Maria H. Daleke, Marcello Ventura, Valentina Donà, Giovanni Delogu, Giorgio Palù, Wilbert Bitter, Riccardo Manganelli

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Abstract

PE are peculiar exported mycobacterial proteins over-represented in pathogenic mycobacterial species. They are characterized by an N-terminal domain of about 110 amino acids (PE domain) which has been demonstrated to be responsible for their export and localization. In this paper, we characterize the PE domain of PE_PGRS33 (PE Rv1818c), one of the best characterized PE proteins. We constructed several mutated proteins in which portions of the PE domain were deleted or subjected to defined mutations. These proteins were expressed in different mycobacterial species and their localization was characterized. We confirmed that the PE domain is essential for PE_PGRS33 surface localization, and demonstrated that a PE domain lacking its first 30 amino acids loses its function. However, single amino acid substitutions in two regions extremely well conserved within the N-terminal domain of all PE proteins had some effect on the stability of PE_PGRS33, but not on its localization. Using Mycobacterium marinum we could show that the type VII secretion system ESX-5 is essential for PE_PGRS33 export. Moreover, in M. marinum, but not in Mycobacterium bovis BCG and in Mycobacterium tuberculosis, the PE domain of PE_PGRS33 is processed and secreted into the culture medium when expressed in the absence of the PGRS domain. Finally, using chimeric proteins in which different portions of the PE Rv1818c domain were fused to the N-terminus of the green fluorescent protein, we could hypothesize that the first 30 amino acids of the PE domain contain a sequence that allows protein translocation.

Original languageEnglish
Article numbere27713
JournalPLoS One
Volume6
Issue number11
DOIs
Publication statusPublished - Nov 16 2011

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ASJC Scopus subject areas

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

Cite this

Cascioferro, A., Daleke, M. H., Ventura, M., Donà, V., Delogu, G., Palù, G., Bitter, W., & Manganelli, R. (2011). Functional dissection of the pe domain responsible for translocation of PE_PGRS33 across the mycobacterial cell wall. PLoS One, 6(11), [e27713]. https://doi.org/10.1371/journal.pone.0027713