Synchronizing protein transport in the secretory pathway
- PMID: 23208546
- DOI: 10.1002/0471143030.cb1519s57
Synchronizing protein transport in the secretory pathway
Abstract
To be secreted or transported to their target compartments, newly synthesized proteins leave the endoplasmic reticulum to reach the Golgi apparatus, where they are processed and sorted toward their final destinations along the secretory pathway. It is now clear that many Golgi-intersecting and non-intersecting pathways exist in cells to carry out proper transport, modification, and addressing. To analyze and visualize the intracellular trafficking of any secretory protein, we developed the retention using selective hooks (RUSH) system. This assay allows the simultaneous release of a pool of particular secretory proteins from the endoplasmic reticulum and the monitoring of their anterograde trafficking. The use of the RUSH system is detailed in these protocols, from sub-cloning the sequence coding for the protein of interest into RUSH plasmids to visualization of its trafficking.
References
Literature Cited
-
- Bard, F., Casano, L., Mallabiabarrena, A., Wallace, E., Saito, K., Kitayama, H., Guizzunti, G., Hu, Y., Wendler, F., Dasgupta, R., Perrimon, N., and Malhotra, V. 2006. Functional genomics reveals genes involved in protein secretion and Golgi organization. Nature 439:604-607.
-
- Boncompain, G., Divoux, S., Gareil, N., de Forges, H., Lescure, A., Latreche, L., Mercanti, V., Jollivet, F., Raposo, G., and Perez, F. 2012. Synchronization of secretory protein traffic in populations of cells. Nat. Methods 9:493-498.
-
- D'Angelo, G., Prencipe, L., Iodice, L., Beznoussenko, G., Savarese, M., Marra, P., Di Tullio, G., Martire, G., De Matteis, M.A., and Bonatti, S. 2009. GRASP65 and GRASP55 sequentially promote the transport of C-terminal valine-bearing cargos to and through the Golgi complex. J. Biol. Chem. 284:34849-34860.
-
- Gordon, D.E., Bond, L.M., Sahlender, D.A., and Peden, A.A. 2010. A targeted siRNA screen to identify SNAREs required for constitutive secretion in mammalian cells. Traffic 11:1191-1204.
-
- Hicks, S.W., Horn, T.A., McCaffery, J.M., Zuckerman, D.M., and Machamer, C.E. 2006. Golgin-160 promotes cell surface expression of the beta-1 adrenergic receptor. Traffic 7:1666-1677.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources