Acidocalcisome-Mitochondrion Membrane Contact Sites in Trypanosoma brucei
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Culture Methods
4.2. Plasmids, Chemicals and Reagents
4.3. Preparation of Guinea Pig Antibody against T. brucei Vacuolar Pyrophosphatase (TbVP1)
4.4. Cell Transfection
4.5. Immunofluorescence Analyses
4.6. Transmission Electron Microscopy
4.7. Proximity Ligation Assay
4.8. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Nolan, D.P.; Voorheis, H.P. The mitochondrion in bloodstream forms of Trypanosoma brucei is energized by the electrogenic pumping of protons catalysed by the F1F0-ATPase. Eur. J. Biochem. 1992, 209, 207–216. [Google Scholar] [CrossRef] [PubMed]
- Vercesi, A.E.; Docampo, R.; Moreno, S.N. Energization-dependent Ca2+ accumulation in Trypanosoma brucei bloodstream and procyclic trypomastigotes mitochondria. Mol. Biochem. Parasitol. 1992, 56, 251–257. [Google Scholar] [CrossRef]
- Schnaufer, A.; Clark-Walker, G.D.; Steinberg, A.G.; Stuart, K. The F1-ATP synthase complex in bloodstream stage trypanosomes has an unusual and essential function. EMBO J. 2005, 24, 4029–4040. [Google Scholar] [CrossRef] [PubMed]
- Brown, S.V.; Hosking, P.; Li, J.; Williams, N. ATP synthase is responsible for maintaining mitochondrial membrane potential in bloodstream form Trypanosoma brucei. Eukaryot. Cell 2006, 5, 45–53. [Google Scholar] [CrossRef] [PubMed]
- Cardenas, C.; Miller, R.A.; Smith, I.; Bui, T.; Molgo, J.; Muller, M.; Vais, H.; Cheung, K.H.; Yang, J.; Parker, I.; et al. Essential regulation of cell bioenergetics by constitutive InsP3 receptor Ca2+ transfer to mitochondria. Cell 2010, 142, 270–283. [Google Scholar] [CrossRef] [PubMed]
- Rizzuto, R.; Brini, M.; Murgia, M.; Pozzan, T. Microdomains with high Ca2+ close to IP3-sensitive channels that are sensed by neighboring mitochondria. Science 1993, 262, 744–747. [Google Scholar] [CrossRef] [PubMed]
- Rizzuto, R.; Pinton, P.; Carrington, W.; Fay, F.S.; Fogarty, K.E.; Lifshitz, L.M.; Tuft, R.A.; Pozzan, T. Close contacts with the endoplasmic reticulum as determinants of mitochondrial Ca2+ responses. Science 1998, 280, 1763–1766. [Google Scholar] [CrossRef] [PubMed]
- Csordas, G.; Thomas, A.P.; Hajnoczky, G. Quasi-synaptic calcium signal transmission between endoplasmic reticulum and mitochondria. EMBO J. 1999, 18, 96–108. [Google Scholar] [CrossRef] [PubMed]
- Csordas, G.; Varnai, P.; Golenar, T.; Roy, S.; Purkins, G.; Schneider, T.G.; Balla, T.; Hajnoczky, G. Imaging interorganelle contacts and local calcium dynamics at the ER-mitochondrial interface. Mol. Cell 2010, 39, 121–132. [Google Scholar] [CrossRef] [PubMed]
- Baughman, J.M.; Perocchi, F.; Girgis, H.S.; Plovanich, M.; Belcher-Timme, C.A.; Sancak, Y.; Bao, X.R.; Strittmatter, L.; Goldberger, O.; Bogorad, R.L.; et al. Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 2011, 476, 341–345. [Google Scholar] [CrossRef] [PubMed]
- De Stefani, D.; Raffaello, A.; Teardo, E.; Szabo, I.; Rizzuto, R. A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 2011, 476, 336–340. [Google Scholar] [CrossRef] [PubMed]
- Huang, G.; Bartlett, P.J.; Thomas, A.P.; Moreno, S.N.; Docampo, R. Acidocalcisomes of Trypanosoma brucei have an inositol 1,4,5-trisphosphate receptor that is required for growth and infectivity. Proc. Natl. Acad. Sci. USA 2013, 110, 1887–1892. [Google Scholar] [CrossRef] [PubMed]
- Lander, N.; Chiurillo, M.A.; Storey, M.; Vercesi, A.E.; Docampo, R. CRISPR/Cas9-mediated endogenous C-terminal tagging of Trypanosoma cruzi genes reveals the acidocalcisome localization of the inositol 1,4,5-trisphosphate receptor. J. Biol. Chem. 2016, 291, 25505–25515. [Google Scholar] [CrossRef] [PubMed]
- Docampo, R.; Huang, G. Calcium signaling in trypanosomatid parasites. Cell Calcium 2015, 57, 194–202. [Google Scholar] [CrossRef] [PubMed]
- Xiong, Z.H.; Ridgley, E.L.; Enis, D.; Olness, F.; Ruben, L. Selective transfer of calcium from an acidic compartment to the mitochondrion of Trypanosoma brucei. Measurements with targeted aequorins. J. Biol. Chem. 1997, 272, 31022–31028. [Google Scholar] [CrossRef] [PubMed]
- Docampo, R.; Lukes, J. Trypanosomes and the solution to a 50-year mitochondrial calcium mystery. Trends Parasitol. 2012, 28, 31–37. [Google Scholar] [CrossRef] [PubMed]
- Tubbs, E.; Rieusset, J. Study of endoplasmic reticulum and mitochondria interactions by in situ proximity ligation assay in fixed cells. J. Vis. Exp. 2016, 118, e54899. [Google Scholar] [CrossRef] [PubMed]
- Tubbs, E.; Theurey, P.; Vial, G.; Bendridi, N.; Bravard, A.; Chauvin, M.A.; Ji-Cao, J.; Zoulim, F.; Bartosch, B.; Ovize, M.; et al. Mitochondria-associated endoplasmic reticulum membrane (MAM) integrity is required for insulin signaling and is implicated in hepatic insulin resistance. Diabetes 2014, 63, 3279–3294. [Google Scholar] [CrossRef] [PubMed]
- Gomez-Suaga, P.; Paillusson, S.; Stoica, R.; Noble, W.; Hanger, D.P.; Miller, C.C. The ER-mitochondria tethering complex VAPB-PTPIP51 regulates autophagy. Curr. Biol. 2017, 27, 371–385. [Google Scholar] [CrossRef] [PubMed]
- Singha, U.K.; Sharma, S.; Chaudhuri, M. Downregulation of mitochondrial porin inhibits cell growth and alters respiratory phenotype in Trypanosoma brucei. Eukaryot. Cell 2009, 8, 1418–1428. [Google Scholar] [CrossRef] [PubMed]
- Chiurillo, M.A.; Lander, N.; Bertolini, M.S.; Storey, M.; Vercesi, A.E.; Docampo, R. Different roles of mitochondrial calcium uniporter complex subunits in growth and infectivity of Trypanosoma cruzi. MBio 2017, 8, e005474-17. [Google Scholar] [CrossRef] [PubMed]
- Lemercier, G.; Dutoya, S.; Luo, S.; Ruiz, F.A.; Rodrigues, C.O.; Baltz, T.; Docampo, R.; Bakalara, N. A vacuolar-type H+-pyrophosphatase governs maintenance of functional acidocalcisomes and growth of the insect and mammalian forms of Trypanosoma brucei. J. Biol. Chem. 2002, 277, 37369–37376. [Google Scholar] [CrossRef] [PubMed]
- Huang, G.; Vercesi, A.E.; Docampo, R. Essential regulation of cell bioenergetics in Trypanosoma brucei by the mitochondrial calcium uniporter. Nat. Commun. 2013, 4, 2865. [Google Scholar] [CrossRef] [PubMed]
- Dominguez, D.C. Calcium signalling in bacteria. Mol. Microbiol. 2004, 54, 291–297. [Google Scholar] [CrossRef] [PubMed]
- Tuteja, N.; Mahajan, S. Calcium signaling network in plants: An overview. Plant Signal. Behav. 2007, 2, 79–85. [Google Scholar] [CrossRef] [PubMed]
- Moreno, S.N.; Docampo, R. Calcium regulation in protozoan parasites. Curr. Opin. Microbiol. 2003, 6, 359–364. [Google Scholar] [CrossRef]
- Clapham, D.E. Calcium signaling. Cell 2007, 131, 1047–1058. [Google Scholar] [CrossRef] [PubMed]
- Burgoyne, T.; Patel, S.; Eden, E.R. Calcium signaling at ER membrane contact sites. Biochim. Biophys. Acta 2015, 1853, 2012–2017. [Google Scholar] [CrossRef] [PubMed]
- King-Keller, S.; Moore, C.A.; Docampo, R.; Moreno, S.N. Ca2+ Regulation of Trypanosoma brucei phosphoinositide phospholipase C. Eukaryot. Cell 2015, 14, 486–494. [Google Scholar] [CrossRef] [PubMed]
- Eisenberg-Bord, M.; Shai, N.; Schuldiner, M.; Bohnert, M. A tether is a tether is a tether: Tethering at membrane contact sites. Dev. Cell 2016, 39, 395–409. [Google Scholar] [CrossRef] [PubMed]
- Kornmann, B.; Currie, E.; Collins, S.R.; Schuldiner, M.; Nunnari, J.; Weissman, J.S.; Walter, P. An ER-mitochondria tethering complex revealed by a synthetic biology screen. Science 2009, 325, 477–481. [Google Scholar] [CrossRef] [PubMed]
- Hirabayashi, Y.; Kwon, S.K.; Paek, H.; Pernice, W.M.; Paul, M.A.; Lee, J.; Erfani, P.; Raczkowski, A.; Petrey, D.S.; Pon, L.A.; et al. ER-mitochondria tethering by PDZD8 regulates Ca2+ dynamics in mammalian neurons. Science 2017, 358, 623–630. [Google Scholar] [CrossRef] [PubMed]
- Wideman, J.G.; Gawryluk, R.M.; Gray, M.W.; Dacks, J.B. The ancient and widespread nature of the ER-mitochondria encounter structure. Mol. Biol. Evol. 2013, 30, 2044–2049. [Google Scholar] [CrossRef] [PubMed]
- Szabadkai, G.; Bianchi, K.; Varnai, P.; De Stefani, D.; Wieckowski, M.R.; Cavagna, D.; Nagy, A.I.; Balla, T.; Rizzuto, R. Chaperone-mediated coupling of endoplasmic reticulum and mitochondrial Ca2+ channels. J. Cell Biol. 2006, 175, 901–911. [Google Scholar] [CrossRef] [PubMed]
- Murley, A.; Nunnari, J. The emerging network of mitochondria-organelle contacts. Mol. Cell 2016, 61, 648–653. [Google Scholar] [CrossRef] [PubMed]
- Phillips, M.J.; Voeltz, G.K. Structure and function of ER membrane contact sites with other organelles. Nat. Rev. Mol. Cell Biol. 2016, 17, 69–82. [Google Scholar] [CrossRef] [PubMed]
- Elbaz-Alon, Y.; Rosenfeld-Gur, E.; Shinder, V.; Futerman, A.H.; Geiger, T.; Schuldiner, M. A dynamic interface between vacuoles and mitochondria in yeast. Dev. Cell 2014, 30, 95–102. [Google Scholar] [CrossRef] [PubMed]
- Honscher, C.; Mari, M.; Auffarth, K.; Bohnert, M.; Griffith, J.; Geerts, W.; van der Laan, M.; Cabrera, M.; Reggiori, F.; Ungermann, C. Cellular metabolism regulates contact sites between vacuoles and mitochondria. Dev. Cell 2014, 30, 86–94. [Google Scholar] [CrossRef] [PubMed]
- Kvam, E.; Goldfarb, D.S. Structure and function of nucleus-vacuole junctions: Outer-nuclear-membrane targeting of Nvj1p and a role in tryptophan uptake. J. Cell Sci. 2006, 119, 3622–3633. [Google Scholar] [CrossRef] [PubMed]
- Wang, H.; Sreenivasan, U.; Hu, H.; Saladino, A.; Polster, B.M.; Lund, L.M.; Gong, D.W.; Stanley, W.C.; Sztalryd, C. Perilipin 5, a lipid droplet-associated protein, provides physical and metabolic linkage to mitochondria. J. Lipid Res. 2011, 52, 2159–2168. [Google Scholar] [CrossRef] [PubMed]
- Miranda, K.; Benchimol, M.; Docampo, R.; de Souza, W. The fine structure of acidocalcisomes in Trypanosoma cruzi. Parasitol. Res. 2000, 86, 373–384. [Google Scholar] [CrossRef] [PubMed]
- Cunningham, I. New culture medium for maintenance of tsetse tissues and growth of trypanosomatids. J. Protozool. 1977, 24, 325–329. [Google Scholar] [CrossRef] [PubMed]
- Hirumi, H.; Hirumi, K. Continuous cultivation of Trypanosoma brucei blood stream forms in a medium containing a low concentration of serum protein without feeder cell layers. J. Parasitol. 1989, 75, 985–989. [Google Scholar] [CrossRef] [PubMed]
- Cordeiro, C.D.; Saiardi, A.; Docampo, R. The inositol pyrophosphate synthesis pathway in Trypanosoma brucei is linked to polyphosphate synthesis in acidocalcisomes. Mol. Microbiol. 2017, 106, 319–333. [Google Scholar] [CrossRef] [PubMed]
- Huang, G.; Ulrich, P.N.; Storey, M.; Johnson, D.; Tischer, J.; Tovar, J.A.; Moreno, S.N.; Orlando, R.; Docampo, R. Proteomic analysis of the acidocalcisome, an organelle conserved from bacteria to human cells. PLoS Pathog. 2014, 10, e1004555. [Google Scholar] [CrossRef] [PubMed]
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ramakrishnan, S.; Asady, B.; Docampo, R. Acidocalcisome-Mitochondrion Membrane Contact Sites in Trypanosoma brucei. Pathogens 2018, 7, 33. https://doi.org/10.3390/pathogens7020033
Ramakrishnan S, Asady B, Docampo R. Acidocalcisome-Mitochondrion Membrane Contact Sites in Trypanosoma brucei. Pathogens. 2018; 7(2):33. https://doi.org/10.3390/pathogens7020033
Chicago/Turabian StyleRamakrishnan, Srinivasan, Beejan Asady, and Roberto Docampo. 2018. "Acidocalcisome-Mitochondrion Membrane Contact Sites in Trypanosoma brucei" Pathogens 7, no. 2: 33. https://doi.org/10.3390/pathogens7020033