In endolysosomal networks, two hetero-hexameric tethers called HOPS and CORVET are found widely throughout eukaryotes. The unicellular ciliate Tetrahymena thermophila possesses elaborate endolysosomal structures, but curiously both it and related protozoa lack the HOPS tether and several other trafficking proteins, while retaining the related CORVET complex. Here, we show that Tetrahymena encodes multiple paralogs of most CORVET subunits, which assemble into six distinct complexes. Each complex has a unique subunit composition and, significantly, shows unique localization, indicating participation in distinct pathways. One pair of complexes differ by a single subunit (Vps8), but have late endosomal versus recycling endosome locations. While Vps8 subunits are thus prime determinants for targeting and functional specificity, determinants exist on all subunits except Vps11. This unprecedented expansion and diversification of CORVET provides a potent example of tether flexibility, and illustrates how 'backfilling' following secondary losses of trafficking genes can provide a mechanism for evolution of new pathways.This article has an associated First Person interview with the first author of the paper.

Biology Centre Institute of Parasitology Czech Academy of Sciences 37005 Ceske Budejovice Czech Republic

Department of Molecular Genetics and Cell Biology 920 E 58th Street The University of Chicago Chicago IL 60637 USA

School of Life Sciences University of Dundee Dundee DD1 5EH UK

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Adl S. M., Leander B. S., Simpson A. G. B., Archibald J. M., Anderson O. R., Bass D., Bowser S. S., Brugerolle G., Farmer M. A., Karpov S. et al. (2007). Diversity, nomenclature, and taxonomy of protists. PubMed DOI

Adl S. M., Simpson A. G. B., Lane C. E., Lukeš J., Bass D., Bowser S. S., Brown M. W., Burki F., Dunthorn M., Hampl V. et al. (2012). The revised classification of eukaryotes. PubMed DOI PMC

Akematsu T., Fukuda Y., Attiq R. and Pearlman R. E. (2014). Role of class III phosphatidylinositol 3-kinase during programmed nuclear death of Tetrahymena thermophila. PubMed DOI PMC

Allen R. D. (2000). The contractile vacuole and its membrane dynamics. PubMed DOI

Asensio C. S., Sirkis D. W., Maas J. W. Jr, Egami K., To T.-L., Brodsky F. M., Shu X., Cheng Y. and Edwards R. H. (2013). Self-assembly of VPS41 promotes sorting required for biogenesis of the regulated secretory pathway. PubMed DOI PMC

Baker R. W. and Hughson F. M. (2016). Chaperoning SNARE assembly and disassembly. PubMed DOI PMC

Balderhaar H. J. and Ungermann C. (2013). CORVET and HOPS tethering complexes - coordinators of endosome and lysosome fusion. PubMed DOI

Balderhaar H. J., Lachmann J., Yavavli E., Brocker C., Lürick A. and Ungermann C. (2013). The CORVET complex promotes tethering and fusion of Rab5/Vps21-positive membranes. PubMed DOI PMC

Bem D., Smith H., Banushi B., Burden J. J., White I. J., Hanley J., Jeremiah N., Rieux-Laucat F., Bettels R., Ariceta G. et al. (2015). VPS33B regulates protein sorting into and maturation of alpha-granule progenitor organelles in mouse megakaryocytes. PubMed DOI PMC

Bright L. J., Kambesis N., Nelson S. B., Jeong B. and Turkewitz A. P. (2010). Comprehensive analysis reveals dynamic and evolutionary plasticity of Rab GTPases and membrane traffic in Tetrahymena thermophila. PubMed DOI PMC

Briguglio J. S., Kumar S. and Turkewitz A. P. (2013). Lysosomal sorting receptors are essential for secretory granule biogenesis in Tetrahymena. PubMed DOI PMC

Brocker C., Kuhlee A., Gatsogiannis C., Balderhaar H. J., Honscher C., Engelbrecht-Vandre S., Ungermann C. and Raunser S. (2012). Molecular architecture of the multisubunit homotypic fusion and vacuole protein sorting (HOPS) tethering complex. PubMed DOI PMC

Chou H.-T., Dukovski D., Chambers M. G., Reinisch K. M. and Walz T. (2016). CATCHR, HOPS and CORVET tethering complexes share a similar architecture. PubMed DOI PMC

Cox J. and Mann M. (2008). MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. PubMed DOI

Cox J., Hein M. Y., Luber C. A., Paron I., Nagaraj N. and Mann M. (2014). Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. PubMed DOI PMC

Cui Y., Zhao Q., Gao C., Ding Y., Zeng Y., Ueda T., Nakano A. and Jiang L. (2014). Activation of the Rab7 GTPase by the MON1-CCZ1 complex is essential for PVC-to-vacuole trafficking and plant growth in arabidopsis. PubMed DOI PMC

Cullinane A. R., Straatman-Iwanowska A., Zaucker A., Wakabayashi Y., Bruce C. K., Luo G., Rahman F., Gürakan F., Utine E., Özkan T. B. et al. (2010). Mutations in VIPAR cause an arthrogryposis, renal dysfunction and cholestasis syndrome phenotype with defects in epithelial polarization. PubMed DOI PMC

Dacks J. B. and Field M. C. (2007). Evolution of the eukaryotic membrane-trafficking system: origin, tempo and mode. PubMed DOI

Dai J., Lu Y., Wang C., Chen X., Fan X., Gu H., Wu X., Wang K., Gartner T. K., Zheng J. et al. (2016). Vps33b regulates Vwf-positive vesicular trafficking in megakaryocytes. PubMed DOI

Davis M. C., Ward J. G., Herrick G. and Allis C. D. (1992). Programmed nuclear death: apoptotic-like degradation of specific nuclei in conjugating Tetrahymena. PubMed DOI

Delevoye C., Hurbain I., Tenza D., Sibarita J.-B., Uzan-Gafsou S., Ohno H., Geerts W. J. C., Verkleij A. J., Salamero J., Marks M. S. et al. (2009). AP-1 and KIF13A coordinate endosomal sorting and positioning during melanosome biogenesis. PubMed DOI PMC

Delevoye C., Heiligenstein X., Ripoll L., Gilles-Marsens F., Dennis M. K., Linares R. A., Derman L., Gokhale A., Morel E., Faundez V. et al. (2016). BLOC-1 brings together the actin and microtubule cytoskeletons to generate recycling endosomes. PubMed DOI PMC

Dennis M. K., Mantegazza A. R., Snir O. L., Tenza D., Acosta-Ruiz A., Delevoye C., Zorger R., Sitaram A., de Jesus-Rojas W., Ravichandran K. et al. (2015). BLOC-2 targets recycling endosomal tubules to melanosomes for cargo delivery. PubMed DOI PMC

Eisen J. A., Coyne R. S., Wu M., Wu D., Thiagarajan M., Wortman J. R., Badger J. H., Ren Q., Amedeo P., Jones K. M. et al. (2006). Macronuclear genome sequence of the ciliate Tetrahymena thermophila, a model eukaryote. PubMed DOI PMC

Elde N. C., Morgan G., Winey M., Sperling L. and Turkewitz A. P. (2005). Elucidation of clathrin-mediated endocytosis in tetrahymena reveals an evolutionarily convergent recruitment of dynamin. PubMed DOI PMC

Elias M., Brighouse A., Gabernet-Castello C., Field M. C. and Dacks J. B. (2012). Sculpting the endomembrane system in deep time: high resolution phylogenetics of Rab GTPases. PubMed DOI PMC

Epp N. and Ungermann C. (2013). The N-terminal domains of Vps3 and Vps8 are critical for localization and function of the CORVET tethering complex on endosomes. PubMed DOI PMC

Frankel J. (2000). Cell biology of Tetrahymena thermophila. PubMed DOI

Fratti R. A., Jun Y., Merz A. J., Margolis N. and Wickner W. (2004). Interdependent assembly of specific regulatory lipids and membrane fusion proteins into the vertex ring domain of docked vacuoles. PubMed DOI PMC

Gabernet-Castello C., O'Reilly A. J., Dacks J. B. and Field M. C. (2013). Evolution of Tre-2/Bub2/Cdc16 (TBC) Rab GTPase-activating proteins. PubMed DOI PMC

Gerst J. E. (1999). SNAREs and SNARE regulators in membrane fusion and exocytosis. PubMed DOI PMC

Gimmler A., Korn R., de Vargas C., Audic S. and Stoeck T. (2016). The Tara Oceans voyage reveals global diversity and distribution patterns of marine planktonic ciliates. PubMed DOI PMC

Gissen P., Johnson C. A., Morgan N. V., Stapelbroek J. M., Forshew T., Cooper W. N., McKiernan P. J., Klomp L. W., Morris A. A. M., Wraith J. E. et al. (2004). Mutations in VPS33B, encoding a regulator of SNARE-dependent membrane fusion, cause arthrogryposis-renal dysfunction-cholestasis (ARC) syndrome. PubMed DOI

Gissen P., Johnson C. A., Gentle D., Hurst L. D., Doherty A. J., O'Kane C. J., Kelly D. A. and Maher E. R. (2005). Comparative evolutionary analysis of VPS33 homologues: genetic and functional insights. PubMed DOI

Guerrier S., Plattner H., Richardson E., Dacks J. B. and Turkewitz A. P. (2017). An evolutionary balance: conservation vs innovation in ciliate membrane trafficking. PubMed DOI PMC

Guo Z., Johnston W., Kovtun O., Mureev S., Bröcker C., Ungermann C. and Alexandrov K. (2013). Subunit organisation of in vitro reconstituted HOPS and CORVET multisubunit membrane tethering complexes. PubMed DOI PMC

Haddad A., Bowman G. R. and Turkewitz A. P. (2002). A new class of cargo protein in Tetrahymena thermophila dense core secretory granules. PubMed DOI PMC

Hausmann K. (1996).

Ho R. and Stroupe C. (2016). The HOPS/class C Vps complex tethers high-curvature membranes via a direct protein-membrane interaction. PubMed DOI

Horazdovsky B. F., Cowles C. R., Mustol P., Holmes M. and Emr S. D. (1996). A novel RING finger protein, Vps8p, functionally interacts with the small GTPase, Vps21p, to facilitate soluble vacuolar protein localization. PubMed DOI

Hunter M., Scourfield E. J., Emmott E. and Graham S. C. (2017). VPS18 recruits VPS41 to the human HOPS complex via a RING-RING interaction. PubMed DOI PMC

Hunter M. R., Hesketh G. G., Benedyk T. H., Gingras A.-C. and Graham S. C. (2018). Proteomic and Biochemical Comparison of the Cellular Interaction Partners of Human VPS33A and VPS33B. PubMed DOI PMC

Huotari J. and Helenius A. (2011). Endosome maturation. PubMed DOI PMC

Jacobs M. E., DeSouza L. V., Samaranayake H., Pearlman R. E., Siu K. W. M. and Klobutcher L. A. (2006). The Tetrahymena thermophila phagosome proteome. PubMed DOI PMC

Jonker C. T. H., Galmes R., Veenendaal T., ten Brink C., van der Welle R. E. N., Liv N., de Rooij J., Peden A. A., van der Sluijs P., Margadant C. et al. (2018). Vps3 and Vps8 control integrin trafficking from early to recycling endosomes and regulate integrin-dependent functions. PubMed DOI PMC

Kaur H., Sparvoli D., Osakada H., Iwamoto M., Haraguchi T. and Turkewitz A. P. (2017). An endosomal syntaxin and the AP-3 complex are required for formation and maturation of candidate lysosome-related secretory organelles (mucocysts) in Tetrahymena thermophila. PubMed DOI PMC

Kiontke S., Langemeyer L., Kuhlee A., Schuback S., Raunser S., Ungermann C. and Kümmel D. (2017). Architecture and mechanism of the late endosomal Rab7-like Ypt7 guanine nucleotide exchange factor complex Mon1-Ccz1. PubMed DOI PMC

Klinger C. M., Klute M. J. and Dacks J. B. (2013). Comparative genomic analysis of multi-subunit tethering complexes demonstrates an ancient pan-eukaryotic complement and sculpting in Apicomplexa. PubMed DOI PMC

Kuhlee A., Raunser S. and Ungermann C. (2015). Functional homologies in vesicle tethering. PubMed DOI

Kümmel D. and Ungermann C. (2014). Principles of membrane tethering and fusion in endosome and lysosome biogenesis. PubMed DOI

LaCava J., Jiang H. and Rout M. P. (2016). Protein complex affinity capture from cryomilled mammalian cells. PubMed DOI PMC

Liu M.-L. and Yao M.-C. (2012). Role of ATG8 and autophagy in programmed nuclear degradation in Tetrahymena thermophila. PubMed DOI PMC

Lo B., Li L., Gissen P., Christensen H., McKiernan P. J., Ye C., Abdelhaleem M., Hayes J. A., Williams M. D., Chitayat D. et al. (2005). Requirement of VPS33B, a member of the Sec1/Munc18 protein family, in megakaryocyte and platelet alpha-granule biogenesis. PubMed DOI

Lobingier B. T. and Merz A. J. (2012). Sec1/Munc18 protein Vps33 binds to SNARE domains and the quaternary SNARE complex. PubMed DOI PMC

Lobingier B. T., Nickerson D. P., Lo S.-Y. and Merz A. J. (2014). SM proteins Sly1 and Vps33 co-assemble with Sec17 and SNARE complexes to oppose SNARE disassembly by Sec18. PubMed DOI PMC

Lorincz P., Lakatos Z., Varga A., Maruzs T., Simon-Vecsei Z., Darula Z., Benko P., Csordas G., Lippai M., Ando I. et al. (2016). MiniCORVET is a Vps8-containing early endosomal tether in Drosophila. PubMed DOI PMC

Lürick A., Gao J., Kuhlee A., Yavavli E., Langemeyer L., Perz A., Raunser S. and Ungermann C. (2017). Multivalent Rab interactions determine tether-mediated membrane fusion. PubMed DOI PMC

Lynch M., Field M. C., Goodson H. V., Malik H. S., Pereira-Leal J. B., Roos D. S., Turkewitz A. P. and Sazer S. (2014). Evolutionary cell biology: two origins, one objective. PubMed DOI PMC

Markgraf D. F., Ahnert F., Arlt H., Mari M., Peplowska K., Epp N., Griffith J., Reggiori F. and Ungermann C. (2009). The CORVET subunit Vps8 cooperates with the Rab5 homolog Vps21 to induce clustering of late endosomal compartments. PubMed DOI PMC

Mellman I. and Yarden Y. (2013). Endocytosis and cancer. PubMed DOI PMC

Miao W., Xiong J., Bowen J., Wang W., Liu Y., Braguinets O., Grigull J., Pearlman R. E., Orias E. and Gorovsky M. A. (2009). Microarray analyses of gene expression during the Tetrahymena thermophila life cycle. PubMed DOI PMC

Morris C., Foster O. K., Handa S., Peloza K., Voss L., Somhegyi H., Jian Y., Vo M. V., Harp M., Rambo F. M. et al. (2018). Function and regulation of the Caenorhabditis elegans Rab32 family member GLO-1 in lysosome-related organelle biogenesis. PubMed DOI PMC

Neefjes J. and van der Kant R. (2014). Stuck in traffic: an emerging theme in diseases of the nervous system. PubMed DOI

Nickerson D. P., Brett C. L. and Merz A. J. (2009). Vps-C complexes: gatekeepers of endolysosomal traffic. PubMed DOI PMC

Nilsson J. R. (1979). Phagotrophy in tetrahymena. In

Nilsson J. R. and Van Deurs B. (1983). Coated pits and pinocytosis in Tetrahymena. PubMed

Nordmann M., Cabrera M., Perz A., Bröcker C., Ostrowicz C., Engelbrecht-Vandré S. and Ungermann C. (2010). The Mon1-Ccz1 complex is the GEF of the late endosomal Rab7 homolog Ypt7. PubMed DOI

Obado S. O., Field M. C., Chait B. T. and Rout M. P. (2016). High-efficiency isolation of nuclear envelope protein complexes from trypanosomes. PubMed DOI

Oeffinger M., Wei K. E., Rogers R., DeGrasse J. A., Chait B. T., Aitchison J. D. and Rout M. P. (2007). Comprehensive analysis of diverse ribonucleoprotein complexes. PubMed DOI

Orias E., Cervantes M. D. and Hamilton E. P. (2011). Tetrahymena thermophila, a unicellular eukaryote with separate germline and somatic genomes. PubMed DOI PMC

Orr A., Song H., Rusin S. F., Kettenbach A. N. and Wickner W. (2017). HOPS catalyzes the interdependent assembly of each vacuolar SNARE into a SNARE complex. PubMed DOI PMC

Ostrowicz C. W., Bröcker C., Ahnert F., Nordmann M., Lachmann J., Peplowska K., Perz A., Auffarth K., Engelbrecht-Vandré S. and Ungermann C. (2010). Defined subunit arrangement and rab interactions are required for functionality of the HOPS tethering complex. PubMed DOI

Peplowska K., Markgraf D. F., Ostrowicz C. W., Bange G. and Ungermann C. (2007). The CORVET tethering complex interacts with the yeast Rab5 homolog Vps21 and is involved in endo-lysosomal biogenesis. PubMed DOI

Perini E. D., Schaefer R., Stöter M., Kalaidzidis Y. and Zerial M. (2014). Mammalian CORVET is required for fusion and conversion of distinct early endosome subpopulations. PubMed DOI

Plattner H. (2010). Membrane trafficking in protozoa SNARE proteins, H+-ATPase, actin, and other key players in ciliates. PubMed DOI

Plattner H. (2015). The contractile vacuole complex of protists--new cues to function and biogenesis. PubMed DOI

Plemel R. L., Lobingier B. T., Brett C. L., Angers C. G., Nickerson D. P., Paulsel A., Sprague D. and Merz A. J. (2011). Subunit organization and Rab interactions of Vps-C protein complexes that control endolysosomal membrane traffic. PubMed DOI PMC

Pulipparacharuvil S., Akbar M. A., Ray S., Sevrioukov E. A., Haberman A. S., Rohrer J. and Kramer H. (2005). Drosophila Vps16A is required for trafficking to lysosomes and biogenesis of pigment granules. PubMed DOI

Rogerson C. and Gissen P. (2018). VPS33B and VIPAR are essential for epidermal lamellar body biogenesis and function. PubMed DOI PMC

Saito-Nakano Y., Nakahara T., Nakano K., Nozaki T. and Numata O. (2010). Marked amplification and diversification of products of ras genes from rat brain, Rab GTPases, in the ciliates Tetrahymena thermophila and Paramecium tetraurelia. PubMed DOI

Schindelin J., Arganda-Carreras I., Frise E., Kaynig V., Longair M., Pietzsch T., Preibisch S., Rueden C., Saalfeld S., Schmid B. et al. (2012). Fiji: an open-source platform for biological-image analysis. PubMed DOI PMC

Schwartz M. L., Nickerson D. P., Lobingier B. T., Plemel R. L., Duan M., Angers C. G., Zick M. and Merz A. J. (2017). Sec17 (alpha-SNAP) and an SM-tethering complex regulate the outcome of SNARE zippering in vitro and in vivo. PubMed DOI PMC

Solinger J. A. and Spang A. (2013). Tethering complexes in the endocytic pathway: CORVET and HOPS. PubMed DOI

Solinger J. A. and Spang A. (2014). Loss of the Sec1/Munc18-family proteins VPS-33.2 and VPS-33.1 bypasses a block in endosome maturation in Caenorhabditis elegans. PubMed DOI PMC

Spang A. (2016). Membrane tethering complexes in the endosomal system. PubMed DOI PMC

Sparvoli D., Richardson E., Osakada H., Lan X., Iwamoto M., Bowman G. R., Kontur C., Bourland W. A., Lynn D. H., Pritchard J. K. et al. (2018). Remodeling the specificity of an endosomal CORVET tether underlies formation of regulated secretory vesicles in the ciliate tetrahymena thermophila. PubMed DOI PMC

Stover N. A., Krieger C. J., Binkley G., Dong Q., Fisk D. G., Nash R., Sethuraman A., Weng S. and Cherry J. M. (2006). Tetrahymena genome database (TGD): a new genomic resource for Tetrahymena thermophila research. PubMed DOI PMC

Strack R. L., Strongin D. E., Bhattacharyya D., Tao W., Berman A., Broxmeyer H. E., Keenan R. J. and Glick B. S. (2008). A noncytotoxic DsRed variant for whole-cell labeling. PubMed DOI PMC

Stroupe C., Collins K. M., Fratti R. A. and Wickner W. (2006). Purification of active HOPS complex reveals its affinities for phosphoinositides and the SNARE Vam7p. PubMed DOI PMC

Takemoto K., Ebine K., Askani J. C., Krüger F., Gonzalez Z. A., Ito E., Goh T., Schumacher K., Nakano A. and Ueda T. (2018). Distinct sets of tethering complexes, SNARE complexes, and Rab GTPases mediate membrane fusion at the vacuole in Arabidopsis. PubMed DOI PMC

Tornieri K., Zlatic S. A., Mullin A. P., Werner E., Harrison R., L'Hernault S. W. and Faundez V. (2013). Vps33b pathogenic mutations preferentially affect VIPAS39/SPE-39-positive endosomes. PubMed DOI PMC

Tyanova S., Temu T., Sinitcyn P., Carlson A., Hein M. Y., Geiger T., Mann M. and Cox J. (2016). The Perseus computational platform for comprehensive analysis of (prote)omics data. PubMed DOI

van der Beek J., Jonker C., van der Welle R., Liv N. and Klumperman J. (2019). CORVET, CHEVI and HOPS - multisubunit tethers of the endo-lysosomal system in health and disease. PubMed DOI

Vizcaíno J. A., Csordas A., del-Toro N., Dianes J. A., Griss J., Lavidas I., Mayer G., Perez-Riverol Y., Reisinger F., Ternent T. et al. (2016). 2016 update of the PRIDE database and its related tools. PubMed DOI PMC

Wang Y., Wang Y., Sheng Y., Huang J., Chen X., Al-Rasheid K. A. S. and Gao S. (2017). A comparative study of genome organization and epigenetic mechanisms in model ciliates, with an emphasis on Tetrahymena, Paramecium and Oxytricha. PubMed DOI

Warren A., Patterson D. J., Dunthorn M., Clamp J. C., Achilles-Day U. E. M., Aescht E., Al-Farraj S. A., Al-Quraishy S., Al-Rasheid K., Carr M. et al. (2017). Beyond the “Code”: a guide to the description and documentation of biodiversity in ciliated protists (Alveolata, Ciliophora). PubMed DOI PMC

Weisse T. (2017). Functional diversity of aquatic ciliates. PubMed DOI

Wurmser A. E., Sato T. K. and Emr S. D. (2000). New component of the vacuolar class C-Vps complex couples nucleotide exchange on the Ypt7 GTPase to SNARE-dependent docking and fusion. PubMed DOI PMC

Xiong J., Lu X., Lu Y., Zeng H., Yuan D., Feng L., Chang Y., Bowen J., Gorovsky M., Fu C. et al. (2011a). Tetrahymena gene expression database (TGED): a resource of microarray data and co-expression analyses for Tetrahymena. PubMed DOI

Xiong J., Yuan D., Fillingham J. S., Garg J., Lu X., Chang Y., Liu Y., Fu C., Pearlman R. E. and Miao W. (2011b). Gene network landscape of the ciliate Tetrahymena thermophila. PubMed DOI PMC

Xiong J., Lu Y., Feng J., Yuan D., Tian M., Chang Y., Fu C., Wang G., Zeng H. and Miao W. (2013). Tetrahymena functional genomics database (TetraFGD): an integrated resource for Tetrahymena functional genomics. PubMed DOI PMC

Zingel P., Agasild H., Karus K., Buholce L. and Nõges T. (2019). Importance of ciliates as food for fish larvae in a shallow sea bay and a large shallow lake. PubMed DOI

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