title: The Sec61 complex is located in both the ER and the ER-Golgi intermediate compartment.
abstract: The heteromeric Sec61 complex is composed of (alpha), beta and (gamma) subunits and forms the core of the mammalian ER translocon. Oligomers of the Sec61 complex form a transmembrane channel where proteins are translocated across and integrated into the ER membrane. We have studied the subcellular localisation of the Sec61 complex using both wild-type COS1 cells and cells transfected with GFP-tagged Sec61(alpha). By double labelling immunofluorescence microscopy the GFP-tagged Sec61(alpha) was found in both the ER and the ER-Golgi intermediate compartment (ERGIC) but not in the trans-Golgi network. Immunofluorescence studies of endogenous Sec61beta and Sec61(gamma) showed that these proteins are also located in both the ER and the ERGIC. Using the alternative strategy of subcellular fractionation, we have shown that wild-type Sec61(alpha), beta and (gamma), and GFP-tagged Sec61(alpha), are all present in both the ER and the ERGIC/Golgi fractions of the gradient. The presence of the Sec61 subunits in a post-ER compartment suggests that these proteins can escape the ER and be recycled back, despite the fact that none of them contain any known membrane protein retrieval signals such as cytosolic di-lysine or di-arginine motifs. We also found that another translocon component, the glycoprotein TRAM, was present in post-ER compartments as demonstrated by subcellular fractionation. Our data indicate that the core components of the mammalian ER translocon are not permanently resident in the ER, but rather that they are maintained in the ER by a specific retrieval mechanism.
authors.affiliation: School of Biological Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK.
authors.forename: J J
authors.initials: JJ
authors.lastname: Greenfield
authors.forename: S
authors.initials: S
authors.lastname: High
mesh_terms: D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D000069816:SEC Translocation Channels
chemical_list: D008164:Luminescent Proteins; D008565:Membrane Proteins; D011500:Protein Synthesis Inhibitors; D000069816:SEC Translocation Channels; D049452:Green Fluorescent Proteins; D020126:Brefeldin A