Type VII Secretion Substrates of Pathogenic Mycobacteria Are Processed by a Surface Protease

M.J. Burggraaf, Alexander Speer, A.S. Meijers, Roy Ummels, Astrid M. van der Sar, K.V. Korotkov, Wilbert Bitter

Research output: Contribution to JournalArticleAcademicpeer-review


Tuberculosis, one of the world’s most severe infectious diseases, is caused by Mycobacterium tuberculosis. A major weapon of this pathogen is a unique cell wall that protects the pathogen from eradication by the immune system. Mycobacteria have specialized secretion systems, e.g., type VII secretion or ESX systems, to transport substrates across this cell wall. The largest group of proteins that are secreted by these ESX systems are the PE proteins. Previously, it was shown that the N-terminal PE domain of about 100 amino acids is required for secretion. Here, we describe the identification of an aspartic protease, designated PecA, that removes (part of) this PE domain at the cell surface. Nearly all of the observed PE_PGRS proteins are processed by PecA. Interestingly, the protease itself is also a secreted PE protein and subject to self-cleavage. Furthermore, a defect in surface processing has no effect on the activity of the PE lipase protein LipY but does seem to affect the functioning of other virulence factors, as a pecA mutant strain of Mycobacterium marinum shows moderate attenuation in zebrafish larvae. In conclusion, our results reveal the presence of a functional aspartic acid protease in M. marinum that cleaves LipY, itself as well as other members of the PE_PGRS family. Finally, mutants lacking PecA show growth attenuation in vivo, suggesting that PecA plays a role during infection. IMPORTANCE Aspartic proteases are common in eukaryotes and retroviruses but are relatively rare among bacteria (N. D. Rawlings and A. Bateman, BMC Genomics 10:437, 2009, https://doi.org/10.1186/1471-2164-10-437). In contrast to eukaryotic aspartic proteases, bacterial aspartic proteases are generally located in the cytoplasm. We have identified a surface-associated mycobacterial aspartic protease, PecA, which cleaves itself and many other type VII secretion substrates of the PE_PGRS family. PecA is present in most pathogenic mycobacterial species, including M. tuberculosis. In addition, pathogenicity of M. marinum is reduced in the ΔpecA mutant, indicating that PecA contributes to virulence.

Original languageEnglish
Article numbere01951-19
Pages (from-to)1-14
Number of pages14
Issue number5
Publication statusPublished - 29 Oct 2019


This work was supported by funding from the National Institute of Allergy and Infectious Diseases (R01AI119022 to K.V.K.), the Cancer Center Amsterdam (to M.J.B. and A.S.M.) and by the Netherlands Organization for Scientific Research (NWO) through a VENI grant (016.Veni.171.090 to A.S.).

FundersFunder number
Cancer Center Amsterdam
Netherlands Organization for Scientific Research
National Institute of Allergy and Infectious DiseasesR01AI119022
Nederlandse Organisatie voor Wetenschappelijk Onderzoek


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