Coated vesicle adaptors
General audience summary:
Cells are divided into special compartments called organelles. But normal cell function requires the constant movement of factors between these compartments, and this is mediated by the cellular transport machinery. Protein and other cargo are packaged into small membrane-bound packages called vesicles for transport, and labelled for delivery to a particular destination. Our research focus is on the role of the ‘adaptor’ proteins that regulate the formation of specific transport vesicles, determining which proteins get bundled into a particular vesicle and where it is targeted to in the cell. Understanding the function and control of adaptor proteins has broad implications in development and in certain diseases such as the hereditary spastic paraplegias that can be caused by mutations in adaptor proteins.
Strategic CIMR themes: Membrane Trafficking, Rare Genetic Diseases
Funding: Wellcome Trust
Research Group members: Anneri Sanger
Research
Proteins are transported between the various organelles of the cell by vesicles, which bud from one membrane and fuse with another. The formation of these vesicles and the selection of the right sort of cargo are dependent on coat proteins. Several types of coated vesicles have been described, the best characterised of which are the clathrin-coated vesicles (CCVs). The coats on CCVs consist primarily of clathrin, adaptor protein (AP) complexes, and 'alternative' adaptors. Our working hypothesis is that for each trafficking pathway, there are a number of different adaptors, each of which is recruited independently onto the appropriate membrane. Once on the membrane, the various adaptors would work together to package different types of cargo into the newly forming vesicle.
We are using several approaches to look for novel adaptors and other components of the trafficking machinery, including proteomic analyses of subcellular fractions, forward genetic screens, and a new method we developed for rapidly inactivating proteins, called 'knocksideways'. Current projects include: establishing the functions of AP-1 and other adaptors in differentiated cells; matching up machinery and cargo proteins; investigating how the WDR11 tethering complex is recruited onto AP-1-derived vesicles; determining why mutations in the non-clathrin adaptors AP-4 and AP-5 cause hereditary spastic paraplegia; and exploring the evolution of adaptors.
Publications
Hirst, J.*, Hesketh, G.G., Gingras, A.-C., and Robinson, M.S.* 2021.
Rag GTPases and phosphatidylinositol 3-phosphate mediate recruitment of the AP-5/SPG11/SPG15 complex. J. Cell Biol. 220(2):e202002075 *Joint corresponding authors
Davies, A.K., Itzhak, D.N., Edgar, J.R., Aechuleta, T.L., Hirst, J., Jackson, L.P., Robinson, M.S.*, and Borner, G.H.H.*. 2018. AP-4 vesicles contribute to spatial control of autophagy via RUSC-dependent peripheral delivery of ATG9A. Nat. Commun. 9: 3958.
*Joint corresponding authors
Hirst J, Itzhak DN, Antrobus R, Borner GHH & Robinson MS. Role of the AP-5 adaptor protein complex in late endosome-to-Golgi retrieval. PLoS Biol. 16: e2004411 (2018).
Navarro Negredo P, Edgar JR, Manna PT, Antrobus R & Robinson MS. 2018. The WDR11 complex facilitates the tethering of AP-1-dervied vesicles. Nature Comm. 9: 596 (2018).
Navarro Negredo P, Edgar JR, Wrobel AG, Zaccai NR, Antrobus R, Owen DJ & Robinson MS. Contribution of the clathrin adaptor AP-1 subunit µ1 to acidic cluster protein sorting. J. Cell Biol. 216, 2927-2943 (2017).
Edgar JR, Manna PT, Nishimura S, Banting G & Robinson MS. Tetherin is an exosomal tether. eLife 5: e17180 (2016).
Hirst J, Edgar JR, Borner GHH, Li S, Sahlender DA, Antrobus R & Robinson MS. Contributions of epsinR and gadkin to clathrin-mediated intracellular trafficking. Mol. Biol. Cell 26, 3085-3103 (2015).
Hirst J, Edgar JR, Esteves T, Darios F, Madeo M, Chang J, Roda RH, Dürr A, Anheim M, Gellera C, Li J, Züchner S, Mariotti C, Stevanin G, Blackstone C, Kruer MC & Robinson MS. Loss of AP-5 results in accumulation of aberrant endolysosomes: defining a new type of lysosomal storage disease. Hum. Mol. Genet. 24, 4984-4996 (2015).
Hirst J, Schlacht A, Norcott JP, Traynor D, Bloomfield G, Antrobus R, Kay RR, Dacks JB & Robinson MS. Characterization of TSET, an ancient and widespread membrane trafficking complex. eLife 3: e02866 (2014).
Borner GHH, Hein MY, Hirst J, Edgar JR, Mann M & Robinson MS. Fractionation profiling: a fast and versatile approach for mapping vesicle proteomics and protein-protein interactions. Mol. Biol. Cell. 25: 3178-3194 (2014).
Kozik P, Hodson NA, Sahlender DA, Simecek N, Soromani C, Wu J, Collinson LM & Robinson MS. A human genome-wide screen for regulators of clathrin-coated vesicle formation reveals an unexpected role for the V-ATPase. Nature Cell Biol. 15, 50–60 (2013).
Borner GHH, Antrobus R, Hirst J, Bhumbra GS, Kozik P, Jackson LP, Sahlender D A & Robinson MS. Multivariate proteomic profiling identifies novel accessory proteins of coated vesicles. J. Cell Biol. 197, 141–160 (2012).
