The p115-interactive proteins GM130 and giantin participate in endoplasmic reticulum-Golgi traffic.
The transport factor p115 is essential for endoplasmic reticulum (ER) to Golgi traffic. P115 interacts with two Golgi proteins, GM130 and giantin, suggesting that they might also participate in ER-Golgi traffic. Here, we show that peptides containing the GM130 or the giantin p115 binding domain and anti-GM130 and anti-giantin antibodies inhibit transport of vesicular stomatitis virus (VSV)-G protein to a mannosidase II-containing Golgi compartment. To determine whether p115, GM130, and giantin act together or sequentially during transport, we compared kinetics of traffic inhibition. Anti-p115, anti-GM130, and anti-giantin antibodies inhibited transport at temporally distinct steps, with the p115-requiring step before the GM130-requiring stage, and both preceding the giantin-requiring stage. Examination of the distribution of the arrested VSV-G protein showed that anti-p115 antibodies inhibited transport at the level of vesicular-tubular clusters, whereas anti-GM130 and anti-giantin antibodies inhibited after the VSV-G protein moved to the Golgi complex. Our results provide the first evidence that GM130 and giantin are required for the delivery of a cargo protein to the mannosidase II-containing Golgi compartment. These data are most consistent with a model where transport from the ER to the cis/medial-Golgi compartments requires the action of p115, GM130, and giantin in a sequential rather than coordinate mechanism.
Alvarez
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Department of Cell Biology, University of Alabama, Birmingham 35294, USA.
Garcia-Mata
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Hauri
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HP
Sztul
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E
D000906:Antibodies / Q000494:pharmacology; D001324:Autoantigens; D002352:Carrier Proteins / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D000077614:Golgi Matrix Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport / Q000187:drug effects; D033921:Vesicular Transport Proteins; D014759:Viral Envelope Proteins / Q000378:metabolism*
D000906:Antibodies; D001324:Autoantigens; D002352:Carrier Proteins; C028391:G protein, vesicular stomatitis virus; D000077614:Golgi Matrix Proteins; C097741:Golgin subfamily A member 2; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010446:Peptide Fragments; D033921:Vesicular Transport Proteins; D014759:Viral Envelope Proteins; C087574:macrogolgin; C091580:vesicular transport factor p115