Group Warren

Golgi Membrane Trafficking Organelle Biogenesis t. Trypanosoma brucei bilobe

Biogenesis of the Golgi apparatus

During normal growth and division, cells double in mass and divide into two equally-sized daughters. All cell constituents must be duplicated and then segregated equally during mitosis. For some constituents, such as the chromosomes, the underlying principles and the mechanistic details are relatively clear. For membrane-bound organelles, such as the Golgi, the principles and mechanism have been controversial. The primary aim of our research work is to understand how the cell creates another copy of the Golgi during the cell cycle and partitions them equally between the two daughter cells, thereby ensuring that this organelle is propagated through successive generations.

The Golgi lies at the heart of the secretory pathway receiving the entire output of newly-synthesized cargo proteins from the endoplasmic reticulum, modifying any bound oligosaccharides, and then sorting them to their final destinations. Typically comprising a stack of closely-apposed and flattened cisternae, the Golgi presents a complex architecture that needs to be duplicated and partitioned every cell cycle. Most studies have focused on the partitioning of the Golgi during mitosis in mammalian cells and most studies suggest that the Golgi undergoes a dramatic conversion to thousands of small vesicles that can then be stochastically distributed between daughter cells. This conversion is triggered by mitotic kinases acting on structural proteins such as GRASPs and golgins.

Golgi duplication has been more difficult to study since most mammalian cells have several hundred copies, subsumed into a ribbon-like structure next to the centrosomes and often the nucleus. This precludes facile observation of the duplication process. We have solved this problem by focusing on organisms that have only a single Golgi that undergoes duplication during the cell cycle and partitioning during mitosis. Protozoan parasites are the best model systems since many have had their genomes sequenced and a variety of molecular biological techniques are available to manipulate protein levels.

Trypanosoma brucei is the causative agent of sleeping sickness in sub-Saharan Africa, and provides a highly-simplified and organized secretory system that is ideal for studying the process of Golgi biogenesis. The duplication of the single Golgi can be observed using GFP-tagged Golgi proteins, and video fluorescence microscopy shows that the old Golgi is involved in the construction of the new. Furthermore, both are located on a novel bilobe structure that appears to act as a template, determining both the size of the Golgi and its inheritance. The composition and duplication of this bilobe are presently under investigation as is the molecular mechanism that generates the new Golgi.

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Morriswood, Brooke; Warren, Graham (2013). Stalemate in the golgi battle. SCIENCE. PMID: 24051247
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Publications since 2006

Kaufmann, Tanja; Kukolj, Eva; Brachner, Andreas; Beltzung, Etienne; Bruno, Melania; Kostrhon, Sebastian; Opravil, Susanne; Hudecz, Otto; Mechtler, Karl; Warren, Graham; Slade, Dea (2016). SIRT2 regulates nuclear envelope reassembly via ANKLE2 deacetylation. J CELL SCI(129): 4607-4621. PMID: 27875273

Morriswood, Brooke; Schmidt, Katy (2015). A MORN Repeat Protein Facilitates Protein Entry into the Flagellar Pocket of Trypanosoma brucei. EUKARYOT CELL;14(11):1081-93. PMID: 26318396

Warren, Graham (2015). In praise of other model organisms. J CELL BIOL;208(4):387-9. PMID: 25688132

McAllaster, Michael R; Ikeda, Kyojiro N; Lozano-Núñez, Ana; Anrather, Dorothea; Unterwurzacher, Verena; Gossenreiter, Thomas; Perry, Jenna A; Crickley, Robbie; Mercadante, Courtney J; Vaughan, Sue; de Graffenried, Christopher L (2015). Proteomic identification of novel cytoskeletal proteins associated with TbPLK, an essential regulator of cell morphogenesis in Trypanosoma brucei. MOL BIOL CELL;26(17):3013-29. PMID: 26133384

Vidilaseris, Keni; Lesigang, Johannes; Morriswood, Brooke; Dong, Gang (2015). Assembly mechanism of Trypanosoma brucei BILBO1 at the flagellar pocket collar. Communicative & integrative biology;8(1):e992739. PMID: 26844754

Vidilaseris, Keni; Morriswood, Brooke; Kontaxis, Georg; Dong, Gang (2014). Structure of the TbBILBO1 N-terminal domain from Trypanosoma brucei reveals an essential requirement for a conserved surface patch. J BIOL CHEM;289(6):3724-35. PMID: 24362019

Sealey-Cardona, Marco; Schmidt, Katy; Demmel, Lars; Hirschmugl, Tatjana; Gesell, Tanja; Dong, Gang; Warren, Graham (2014). Sec16 determines the size and functioning of the Golgi in the protist parasite, Trypanosoma brucei. TRAFFIC;15(6):613-29. PMID: 24612401

Demmel, Lars; Schmidt, Katy; Lucast, Louise; Havlicek, Katharina; Zankel, Armin; Koestler, Tina; Reithofer, Viktoria; de Camilli, Pietro; Warren, Graham (2014). The endocytic activity of the flagellar pocket in Trypanosoma brucei is regulated by an adjacent phosphatidylinositol phosphate kinase. J CELL SCI;127(Pt 10):2351-64. PMID: 24639465

Vidilaseris, Keni; Shimanovskaya, Ekaterina; Esson, Heather J; Morriswood, Brooke; Dong, Gang (2014). Assembly Mechanism of Trypanosoma brucei BILBO1, a Multidomain Cytoskeletal Protein. J BIOL CHEM;289(34):23870-81. PMID: 25031322

Morriswood, Brooke; Havlicek, Katharina; Demmel, Lars; Yavuz, Sevil; Sealey-Cardona, Marco; Vidilaseris, Keni; Anrather, Dorothea; Kostan, Julius; Djinovic-Carugo, Kristina; Roux, Kyle; Warren, Graham (2013). Novel bilobe components in Trypanosoma brucei identified using proximity-dependent biotinylation. EUKARYOT CELL;12(2):356-67. PMID: 23264645

Misteli, Tom; Warren, Graham (2013). 25 Years of Current Opinion in Cell Biology. CURR OPIN CELL BIOL;1(25):1-2. PMID: 23352256

Lozano-Núñez, Ana; Ikeda, Kyojiro N; Sauer, Thomas; de Graffenried, Christopher L (2013). An analogue-sensitive approach identifies basal body rotation and flagellum attachment zone elongation as key functions of PLK in Trypanosoma brucei. MOL BIOL CELL;24(9):1321-33. PMID: 23447704

de Graffenried, Christopher L; Anrather, Dorothea; Von Raußendorf, Freia; Warren, Graham (2013). Polo-like kinase phosphorylation of bilobe-resident TbCentrin2 facilitates flagellar inheritance in Trypanosoma brucei. MOL BIOL CELL;24(12):1947-63. PMID: 23615446

Warren, Graham (2013). Transport through the Golgi in Trypanosoma brucei. HISTOCHEM CELL BIOL;140(3):235-8. PMID: 23765165

Morriswood, Brooke; Warren, Graham (2013). Cell biology. Stalemate in the Golgi battle. SCIENCE;341(6153):1465-6. PMID: 24051247

Warren, Graham (2013). Size and position matter. NAT REV MOL CELL BIO;14(12):755-7. PMID: 24409512

Esson, Heather J; Morriswood, Brooke; Yavuz, Sevil; Vidilaseris, Keni; Dong, Gang; Warren, Graham (2012). Morphology of the trypanosome bilobe, a novel cytoskeletal structure. EUKARYOT CELL;11(6):761-772 . PMID: 22327007

Ikeda, Kyojiro N. and Christopher L. de Graffenried (2012). Polo-like kinase is necessary for flagellum inheritance in Trypanosoma brucei. J CELL SCI;125:3173–3184. PMID: 22427687

Warren, Graham (2012). A vital assay. NAT REV MOL CELL BIO;13(12):754. PMID: 23151658

Demmel, Lars; Melak, Michael; Kotisch, Harald; Fendos, Justin; Reipert, Siegfried; Warren, Graham (2011). Differential Selection of Golgi Proteins by COPII Sec24 Isoforms in Procyclic Trypanosoma brucei. TRAFFIC;11(12):1575-91. PMID: 21801288

Esson, Heather J., Leander, Brian S. (2010). Evolution of Distorted Pellicle Patterns in Rigid Photosynthetic Euglenids (Phacus Dujardin). J EUKARYOT MICROBIOL;57(1):19-32. PMID: 19878404

Rutz, Christoph; Satoh, Ayano; Ronchi, Paolo; Brügger, Britta; Warren, Graham; Wieland, Felix T (2009). Following the fate in vivo of COPI vesicles generated in vitro. TRAFFIC. PMID: 19497049

Morriswood, Brooke; He, Cynthia Y; Sealey-Cardona, Marco; Yelinek, Jordan; Pypaert, Marc; Warren, Graham (2009). The bilobe structure of Trypanosoma brucei contains a MORN-repeat protein. MOL BIOCHEM PARASIT. PMID: 19445968

Yelinek, Jordan T; He, Cynthia Y; Warren, Graham (2009). Ultrastructural study of Golgi duplication in Trypanosoma brucei. TRAFFIC. PMID: 19207482

Scott Emr,Benjamin S. Glick,Adam D.Linstedt,Jennifer Lippincott-Schwartz, Alberto Luini,Vivek Malhotra,Brad J. Marsh, Akihiko Nakano,Suzanne R. Pfeffer,Catherine Rabouille,James E. Rothman,Graham Warren, and Felix T. Wieland (2009). Journeys through the Golgi—taking stock in a new era. J CELL BIOL;187(4):449. PMID: 19948493

Shi, Jie; Franklin, Joseph B; Yelinek, Jordan T; Ebersberger, Ingo; Warren, Graham; He, Cynthia Y (2008). Centrin4 coordinates cell and nuclear division in T. brucei. J CELL SCI. PMID: 18768932

Satoh, Ayano; Warren, Graham (2008). In situ cleavage of the acidic domain from the p115 tether inhibits exocytic transport. TRAFFIC. PMID: 18564369

Tang, Danming; Mar, Kari; Warren, Graham; Wang, Yanzhuang (2008). Molecular mechanism of mitotic Golgi disassembly and reassembly revealed by a defined reconstitution assay. J BIOL CHEM. PMID: 18156178

de Graffenried, Christopher L; Ho, Helen H; Warren, Graham (2008). Polo-like kinase is required for Golgi and bilobe biogenesis in Trypanosoma brucei. J CELL BIOL. PMID: 18443217

Ramirez, Irene Barinaga-Rementeria; de Graffenried, Christopher L; Ebersberger, Ingo; Yelinek, Jordan; He, Cynthia Y; Price, Albert; Warren, Graham (2008). TbG63, a golgin involved in Golgi architecture in Trypanosoma brucei. J CELL SCI. PMID: 18411253

Müller, J M M; Deinhardt, K; Rosewell, I; Warren, G; Shima, D T (2007). Targeted deletion of p97 (VCP/CDC48) in mouse results in early embryonic lethality. BIOCHEM BIOPH RES CO. PMID: 17239345

Gniadek, Thomas J; Warren, Graham (2007). WatershedCounting3D: a new method for segmenting and counting punctate structures from confocal image data. TRAFFIC. PMID: 17319897

Hartmann, Jan; Hu, Ke; He, Cynthia Y; Pelletier, Laurence; Roos, David S; Warren, Graham (2006). Golgi and centrosome cycles in Toxoplasma gondii. MOL BIOCHEM PARASIT. PMID: 16266757

Ho, Helen H; He, Cynthia Y; de Graffenried, Christopher L; Murrells, Lindsay J; Warren, Graham (2006). Ordered assembly of the duplicating Golgi in Trypanosoma brucei. P NATL ACAD SCI USA. PMID: 16672362