Abstract
Rud3p is a coiled-coil protein of the yeast cis-Golgi. We find that Rud3p is localized to the Golgi via a COOH-terminal domain that is distantly related to the GRIP domain that recruits several coiled-coil proteins to the trans-Golgi by binding the small Arf-like GTPase Arl1p. In contrast, Rud3p binds to the GTPase Arf1p via this COOH-terminal "GRIP-related Arf-binding" (GRAB) domain. Deletion of RUD3 is lethal in the absence of the Golgi GTPase Ypt6p, and a screen of other mutants showing a similar genetic interaction revealed that Golgi targeting of Rud3p also requires Erv14p, a cargo receptor that cycles between the endoplasmic reticulum and Golgi. The one human protein with a GRAB domain, GMAP-210 (CEV14/Trip11/Trip230), is known to be on the cis-Golgi, but the COOH-terminal region that contains the GRAB domain has been reported to bind to centrosomes and gamma-tubulin (Rios, R.M, A. Sanchis, A.M. Tassin, C. Fedriani, and M. Bornens. 2004. Cell. 118:323-335). In contrast, we find that this region binds to the Golgi in a GRAB domain-dependent manner, suggesting that GMAP-210 may not link the Golgi to gamma-tubulin and centrosomes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ADP-Ribosylation Factor 1 / metabolism*
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Amino Acid Motifs
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Amino Acid Sequence
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Animals
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COS Cells
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Carrier Proteins / metabolism*
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Chromatography
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Cytoskeletal Proteins
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Endoplasmic Reticulum / metabolism*
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GTP Phosphohydrolases / metabolism*
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Gene Deletion
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Genotype
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Golgi Apparatus / metabolism*
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Guanosine Diphosphate / metabolism
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Membrane Proteins / metabolism*
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Microscopy, Confocal
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Microscopy, Fluorescence
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Microtubule-Associated Proteins / metabolism
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Molecular Sequence Data
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Monomeric GTP-Binding Proteins / metabolism
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Mutation
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Nuclear Proteins
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Plasmids / metabolism
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Protein Conformation
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Protein Structure, Tertiary
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Recombinant Proteins / chemistry
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / metabolism*
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Sequence Homology, Amino Acid
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Tubulin / metabolism
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Vesicular Transport Proteins
Substances
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Carrier Proteins
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Cytoskeletal Proteins
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Erv14 protein, S cerevisiae
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Membrane Proteins
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Microtubule-Associated Proteins
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Nuclear Proteins
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RUD3 protein, S cerevisiae
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Recombinant Proteins
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Saccharomyces cerevisiae Proteins
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TRIP11 protein, human
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Tubulin
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Vesicular Transport Proteins
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Guanosine Diphosphate
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GTP Phosphohydrolases
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ADP-Ribosylation Factor 1
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Monomeric GTP-Binding Proteins
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YPT6 protein, S cerevisiae