GS15 forms a SNARE complex with syntaxin 5, GS28, and Ykt6 and is implicated in traffic in the early cisternae of the Golgi apparatus.
The subcellular localization, interacting partners, and function of GS15, a Golgi SNARE, remain to be established. In our present study, it is revealed that unlike proteins (Bet1 and the KDEL receptor) cycling between the Golgi and the intermediate compartment (IC, inclusive of the ER exit sites), GS15 is not redistributed into the IC upon incubation at 15 degrees C or when cells are treated with brefeldin A. Immuno-electron microscopy (immuno-EM) reveals that GS15 is mainly found in the medial-cisternae of the Golgi apparatus and adjacent tubulo-vesicular elements. Coimmunoprecipitation experiments suggest that GS15 exists in a distinct SNARE complex that contains SNAREs (syntaxin5, GS28, and Ykt6) that are implicated in both ER-to-Golgi and intra-Golgi transport but not with SNAREs involved exclusively in ER-to-Golgi traffic. Furthermore, components of COPI coat can be selectively coimmunoprecipitated with GS15 from Golgi extracts. Overexpression of mutant forms of GS15 affects the normal distribution of cis- and medial-Golgi proteins (GS28, syntaxin 5, and Golgi mannosidase II), whereas proteins of the trans-Golgi and TGN (Vti1-rp2/Vti1a and syntaxin 6) and Golgi matrix/scaffold (GM130 and p115) are less affected. When the level of GS15 is reduced by duplex 21-nt small interfering RNA (siRNA)-mediated knockdown approach, diverse markers of the Golgi apparatus are redistributed into small dotty and diffuse labeling, suggesting an essential role of GS15 in the Golgi apparatus.
Xu
Yue
Y
Membrane Biology Laboratory, Institute of Molecular and Cell Biology, Singapore 117609, Singapore.
Martin
Sally
S
James
David E
DE
Hong
Wanjin
W
D000818:Animals; D020126:Brefeldin A / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D020755:Coat Protein Complex I / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D046911:Macromolecular Substances; D008361:Mannosidases / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D050683:R-SNARE Proteins; D034741:RNA, Small Interfering / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins / Q000378:metabolism
C098783:BET1 protein, rat; C107588:BET1L protein, human; C495951:Bet1l protein, rat; D002352:Carrier Proteins; D020755:Coat Protein Complex I; C490972:GOSR1 protein, human; C064459:KDEL receptor; D046911:Macromolecular Substances; D008565:Membrane Proteins; D011500:Protein Synthesis Inhibitors; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D050683:R-SNARE Proteins; D034741:RNA, Small Interfering; D018000:Receptors, Peptide; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins; C491482:YKT6 protein, human; C491481:Ykt6 protein, rat; D020126:Brefeldin A; D008361:Mannosidases; C038829:mannosyl-oligosaccharide 1,3 - 1,6-alpha-mannosidase