Abstract
Plasmodium falciparum exports hundreds of virulence proteins within infected erythrocytes, a process that requires cleavage of a pentameric motif called Plasmodium export element or vacuolar transport signal by the endoplasmic reticulum (ER)-resident protease plasmepsin V. We identified plasmepsin V-binding proteins that form a unique interactome required for the translocation of effector cargo into the parasite ER. These interactions are functionally distinct from the Sec61–signal peptidase complex required for the translocation of proteins destined for the classical secretory pathway. This interactome does not involve the signal peptidase (SPC21) and consists of PfSec61, PfSPC25, plasmepsin V and PfSec62, which is an essential component of the post-translational ER translocon. Together, they form a distinct portal for the recognition and translocation of a large subset of Plasmodium export element effector proteins into the ER, thereby remodelling the infected erythrocyte that is required for parasite survival and pathogenesis.
Similar content being viewed by others
References
The World Malaria Report (WHO, 2015); http://www.who.int/malaria/publications/world-malaria-report-2015/report/en/.
Cowman, A. F. et al. Malaria: biology and disease. Cell 167, 610–624 (2016).
Maier, A. G. et al. Exported proteins required for virulence and rigidity of Plasmodium falciparum-infected human erythrocytes. Cell 134, 48–61 (2008).
Marti, M. et al. Targeting malaria virulence and remodeling proteins to the host erythrocyte. Science 306, 1930–1933 (2004).
Hiller, N. L. et al. A host-targeting signal in virulence proteins reveals a secretome in malarial infection. Science 306, 1934–1937 (2004).
Sargeant, T. et al. Lineage-specific expansion of proteins exported to erythrocytes in malaria parasites. Genome Biol. 7, R12 (2006).
Chang, H. H. et al. N-terminal processing of proteins exported by malaria parasites. Mol. Biochem. Parasitol. 160, 107–115 (2008).
Boddey, J. A. et al. Role of the Plasmodium export element in trafficking parasite proteins to the infected erythrocyte. Traffic 10, 285–299 (2009).
Boddey, J. A. et al. An aspartyl protease directs malaria effector proteins to the host cell. Nature 463, 627–631 (2010).
Russo, I. et al. Plasmepsin V licenses Plasmodium proteins for export into the host erythrocyte. Nature 463, 632–636 (2010).
Boddey, J. A. et al. Export of malaria proteins requires co-translational processing of the PEXEL motif independent of phosphatidylinositol-3-phosphate binding. Nat. Commun. 7, 10470 (2016).
Sleebs, B. E. et al. Inhibition of plasmepsin V activity demonstrates its essential role in protein export, PfEMP1 display, and survival of malaria parasites. PLoS Biol. 12, e1001897 (2014).
Hodder, A. N. et al. Structural basis for plasmepsin V inhibition that blocks export of malaria proteins to human erythrocytes. Nat. Struct. Mol. Biol. 22, 590–596 (2015).
Przyborski, J.M., Nyboer, B., Lanzer, M. Ticket to ride: Export of proteins to the P. falciparum infected erythrocyte. Mol. Micro. 101, 1–11 (2016).
Gilmore, R. The protein translocation apparatus of the rough endoplasmic reticulum, its associated proteins, and the mechanism of translocation. Curr. Opin. Cell Biol. 3, 580–584 (1991).
Marapana, D. S. et al. Malaria parasite signal peptide peptidase is an ER-resident protease required for growth but not for invasion. Traffic 13, 1457–1465 (2012).
Harbut, M. B. et al. Targeting the ERAD pathway via inhibition of signal peptide peptidase for antiparasitic therapeutic design. Proc. Natl Acad. Sci. USA 109, 21486–21491 (2012).
Shelness, G. S., Kanwar, Y. S. & Blobel, G. cDNA-derived primary structure of the glycoprotein component of canine microsomal signal peptidase complex. J. Biol. Chem. 263, 17063–17070 (1988).
Prommana, P. et al. Inducible knockdown of Plasmodium gene expression using the glmS ribozyme. PLoS ONE 8, e73783 (2013).
Elsworth, B. et al. PTEX is an essential nexus for protein export in malaria parasites. Nature 511, 587–591 (2014).
de Koning-Ward, T. F. et al. A newly discovered protein export machine in malaria parasites. Nature 459, 945–949 (2009).
Riglar, D. T. et al. Spatial association with PTEX complexes defines regions for effector export into Plasmodium falciparum-infected erythrocytes. Nat. Commun. 4, 1415 (2013).
Aikawa, M. et al. Pf155/RESA antigen is localized in dense granules of Plasmodium falciparum merozoites. Exp. Parasitol. 71, 326–329 (1990).
Dixon, M. W. et al. Targeting of the ring exported protein 1 to the Maurer’s clefts is mediated by a two-phase process. Traffic 9, 1316–1326 (2008).
Blisnick, T. et al. Pfsbp1, a Maurer’s cleft Plasmodium falciparum protein, is associated with the erythrocyte skeleton. Mol. Biochem. Parasitol. 111, 107–121 (2000).
Boddey, J. A. & Cowman, A. F. Plasmodium nesting: remaking the erythrocyte from the inside out. Annu. Rev. Microbiol. 67, 243–269 (2013).
Kalies, K. U., Rapoport, T. A. & Hartmann, E. The β subunit of the Sec61 complex facilitates cotranslational protein transport and interacts with the signal peptidase during translocation. J. Cell Biol. 141, 887–894 (1998).
Antonin, W., Meyer, H. A. & Hartmann, E. Interactions between Spc2p and other components of the endoplasmic reticulum translocation sites of the yeast Saccharomyces cerevisiae. J. Biol. Chem. 275, 34068–34072 (2000).
Kalies, K. U. & Hartmann, E. Membrane topology of the 12- and the 25-kDa subunits of the mammalian signal peptidase complex. J. Biol. Chem. 271, 3925–3929 (1996).
Evans, E. A., Gilmore, R. & Blobel, G. Purification of microsomal signal peptidase as a complex. Proc. Natl Acad. Sci. USA 83, 581–585 (1986).
Rothblatt, J. A. et al. Multiple genes are required for proper insertion of secretory proteins into the endoplasmic reticulum in yeast. J. Cell Biol. 109, 2641–2652 (1989).
Ciu, J. et al. Competitive inhibition of the endoplasmic reticulum signal peptidase by non-cleavable mutant preprotein cargos. J. Biol. Chem. 290, 28131–28140 (2015).
Deshaies, R. J. & Schekman, R. SEC62 encodes a putative membrane protein required for protein translocation into the yeast endoplasmic reticulum. J. Cell Biol. 109, 2653–2664 (1989).
Mullins, C. et al. Structurally related Spc1p and Spc2p of yeast signal peptidase complex are functionally distinct. J. Biol. Chem. 271, 29094–29099 (1996).
Nunn, D. N. & Lory, S. Product of the Pseudomonas aeruginosa gene pilD is a prepilin leader peptidase. Proc. Natl Acad. Sci. USA 88, 3281–3285 (1991).
Tokunaga, M. et al. Prolipoprotein signal peptidase in Escherichia coli is distinct from the M13 procoat protein signal peptidase. J. Biol. Chem. 257, 9922–9925 (1982).
Spiller, M. P. & Stirling, C. J. Preferential targeting of a signal recognition particle-dependent precursor to the Ssh1p translocon in yeast. J. Biol. Chem. 286, 21953–21960 (2011).
Feltcher, M. E. & Braunstein, M. Emerging themes in SecA2-mediated protein export. Nat. Rev. Microbiol. 10, 779–789 (2012).
Ghorbal, M. et al. Genome editing in the human malaria parasite Plasmodium falciparum using the CRISPR–Cas9 system. Nat. Biotechnol. 32, 819–821 (2014).
Tatebe, K. Combining multiple averaged data points and their errors (White paper, 2005).
Prieto, D. A., Ye, X. & Veenstra, T. D. Proteomic analysis of traumatic brain injury: the search for biomarkers. Expert Rev. Proteomics 5, 283–291 (2008).
Nesvizhskii, A. I. et al. A statistical model for identifying proteins by tandem mass spectrometry. Anal. Chem. 75, 4646–4658 (2003).
Searle, B. C. Scaffold: a bioinformatic tool for validating MS/MS-based proteomic studies. Proteomics 10, 1265–1269 (2010).
Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367–1372 (2008).
Nguyen, H. D., Wood, I. A. & Hill, M. M. A robust permutation test for quantitative SILAC proteoics experiments. JIOmics 2, 80–93 (2012).
Wisniewski, J. R., Zougman, A. & Mann, M. Combination of FASP and StageTip-based fractionation allows in-depth analysis of the hippocampal membrane proteome. J. Proteome Res. 8, 5674–5678 (2009).
Delconte, R. B. et al. CIS is a potent checkpoint in NK cell-mediated tumor immunity. Nat. Immunol. 17, 816–824 (2016).
Cox, J. et al. Andromeda: a peptide search engine integrated into the MaxQuant environment. J. Proteome Res. 10, 1794–1805 (2011).
Cox, J. et al. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol. Cell. Proteomics 13, 2513–2526 (2014).
Keilhauer, E. C., Hein, M. Y. & Mann, M. Accurate protein complex retrieval by affinity enrichment mass spectrometry (AE-MS) rather than affinity purification mass spectrometry (AP-MS). Mol. Cell. Proteomics 14, 120–135 (2015).
Acknowledgements
We thank the Red Cross Blood Service (Melbourne, Victoria, Australia) for the supply of donor blood for our cell culture. This work was supported by the NHMRC of Australia and the Victorian State Government Operational Infrastructure Support and the Australian Government NHMRC IRIISS. A.F.C. is a Howard Hughes International Scholar. M.P. was supported by the EMBO Long-Term Fellowship ALTF 793-2016.
Author information
Authors and Affiliations
Contributions
D.S.M. conceived and performed the experiments, acquired and analysed the data and wrote the manuscript. L.F.D. conceived and performed the experiments, acquired and analysed the data and edited the manuscript. J.J.S. conceived and performed the experiments, acquired and analysed the data and edited the manuscript. T.N. conceived and performed the experiments, acquired and analysed the data and edited the manuscript. T.T. performed the experiments, acquired and analysed the data and edited the manuscript. M.P. performed the experiments, acquired and analysed the data and edited the manuscript. B.K.D. conceived and performed the experiments, acquired and analysed the data and edited the manuscript. B.S.C. conceived the experiments, analysed the data and edited the manuscript. P.R.G. conceived the experiments, analysed the data and edited the manuscript. A.I.W. conceived and performed the experiments, acquired and analysed the data and edited the manuscript. J.A.B. conceived the experiments, analysed the data and edited the manuscript. A.F.C. conceived the experiments, analysed the data and wrote the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Supplementary Information
Supplementary Figures 1–13, Supplementary Table 2 and Supplementary Table 5.
Rights and permissions
About this article
Cite this article
Marapana, D.S., Dagley, L.F., Sandow, J.J. et al. Plasmepsin V cleaves malaria effector proteins in a distinct endoplasmic reticulum translocation interactome for export to the erythrocyte. Nat Microbiol 3, 1010–1022 (2018). https://doi.org/10.1038/s41564-018-0219-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41564-018-0219-2
- Springer Nature Limited
This article is cited by
-
Toxoplasma protein export and effector function
Nature Microbiology (2024)
-
Aryl amino acetamides prevent Plasmodium falciparum ring development via targeting the lipid-transfer protein PfSTART1
Nature Communications (2024)
-
Tryptophan C-mannosylation is critical for Plasmodium falciparum transmission
Nature Communications (2022)
-
A cell surface-exposed protein complex with an essential virulence function in Ustilago maydis
Nature Microbiology (2021)
-
The parasitophorous vacuole of the blood-stage malaria parasite
Nature Reviews Microbiology (2020)