Limits...
The GTPase Arf1p and the ER to Golgi cargo receptor Erv14p cooperate to recruit the golgin Rud3p to the cis-Golgi.

Gillingham AK, Tong AH, Boone C, Munro S - J. Cell Biol. (2004)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, England, UK.

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.

Show MeSH

Related in: MedlinePlus

Rud3p is a member of a family of coiled-coil proteins with a conserved COOH-terminal domain. (A) Schematic representation of S. cerevisiae Rud3p and its relatives from the indicated species. At, A. thaliana; Ce, C. elegans; Dm, D. melanogaster; Hs, Homo sapiens; Sc, S. cerevisiae; Tb, Trypanosoma brucei. (B) Alignment of the COOH-terminal regions of the GRAB domain proteins with those of GRIP domain proteins golgin-245 and golgin-97, and the structure of the GRIP domain of human golgin-245. The two sets of sequences were independently aligned with CLUSTAL W and shaded where more than half the residues are related (gray) or identical (black). Hydrophobic residues conserved in each set are marked with filled circles and with a red circle for the critical tyrosine in the GRIP domain, and the leucine is in the equivalent position in Rud3p. In both alignments the tryptophans are shaded orange and cluster downstream of the conserved region. In the case of golgin-245, the tryptophan apparently stabilizes the interaction of the GRIP domain with Golgi membranes (Panic et al., 2003a). (C) A schematic representation of the GRAB domain proteins from metazoans and yeasts, along with a GRIP domain protein. All contain either a GRIP or GRAB domain, and the latter have a downstream GA1 motif. Metazoan GRAB proteins also have an extended, proline-rich, COOH-terminal region, whereas GRAB proteins from yeasts and filamentous fungi have an upstream region of sequence conservation (GA2, blue). (D) Fluorescent micrographs of rud3Δ cells expressing GFP-tagged wild-type Rud3p or the mutant L410A as in Fig. 1 A.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2172552&req=5

fig2: Rud3p is a member of a family of coiled-coil proteins with a conserved COOH-terminal domain. (A) Schematic representation of S. cerevisiae Rud3p and its relatives from the indicated species. At, A. thaliana; Ce, C. elegans; Dm, D. melanogaster; Hs, Homo sapiens; Sc, S. cerevisiae; Tb, Trypanosoma brucei. (B) Alignment of the COOH-terminal regions of the GRAB domain proteins with those of GRIP domain proteins golgin-245 and golgin-97, and the structure of the GRIP domain of human golgin-245. The two sets of sequences were independently aligned with CLUSTAL W and shaded where more than half the residues are related (gray) or identical (black). Hydrophobic residues conserved in each set are marked with filled circles and with a red circle for the critical tyrosine in the GRIP domain, and the leucine is in the equivalent position in Rud3p. In both alignments the tryptophans are shaded orange and cluster downstream of the conserved region. In the case of golgin-245, the tryptophan apparently stabilizes the interaction of the GRIP domain with Golgi membranes (Panic et al., 2003a). (C) A schematic representation of the GRAB domain proteins from metazoans and yeasts, along with a GRIP domain protein. All contain either a GRIP or GRAB domain, and the latter have a downstream GA1 motif. Metazoan GRAB proteins also have an extended, proline-rich, COOH-terminal region, whereas GRAB proteins from yeasts and filamentous fungi have an upstream region of sequence conservation (GA2, blue). (D) Fluorescent micrographs of rud3Δ cells expressing GFP-tagged wild-type Rud3p or the mutant L410A as in Fig. 1 A.

Mentions: The majority of Rud3p is predicted to form a coiled-coil (Fig. 1 B), but the last 80 residues that mediate targeting lie outside this region. When this COOH-terminal region was used to search the GenBank database using the iterative program PSI-BLAST, it revealed that a large coiled-coil protein with a COOH-terminal domain related to the COOH-terminal 80 residues of Rud3p is present in the genomes of most eukaryotes including mammals, Drosophila melanogaster, Caenorhabditis elegans, plants, fungi, and Dictyostelium discoideum (Fig. 2, A and B). In all but one of these species there is a single member of the family present, the exception being Arabidopsis thaliana, which has two such proteins that are very closely related. The only one of these proteins from other species to have been previously characterized is that from humans, which is called GMAP-210, CEV14, Trip11, or Trip230. This protein was independently identified by either screening an expression library with a human auto-antiserum (GMAP-210, p210; Rios et al., 1994) or by yeast two-hybrid screens with thyroid receptor (Trip11; Lee et al., 1995) and retinoblastoma protein (Trip230; Chang et al., 1997), or as a gene found fused to that of the platelet-derived growth factor receptor by an oncogenic chromosomal translocation (CEV14; Abe et al., 1997). Immunoelectron microscopy has shown that GMAP-210 is localized to the cis side of the Golgi apparatus (Rios et al., 1994).


The GTPase Arf1p and the ER to Golgi cargo receptor Erv14p cooperate to recruit the golgin Rud3p to the cis-Golgi.

Gillingham AK, Tong AH, Boone C, Munro S - J. Cell Biol. (2004)

Rud3p is a member of a family of coiled-coil proteins with a conserved COOH-terminal domain. (A) Schematic representation of S. cerevisiae Rud3p and its relatives from the indicated species. At, A. thaliana; Ce, C. elegans; Dm, D. melanogaster; Hs, Homo sapiens; Sc, S. cerevisiae; Tb, Trypanosoma brucei. (B) Alignment of the COOH-terminal regions of the GRAB domain proteins with those of GRIP domain proteins golgin-245 and golgin-97, and the structure of the GRIP domain of human golgin-245. The two sets of sequences were independently aligned with CLUSTAL W and shaded where more than half the residues are related (gray) or identical (black). Hydrophobic residues conserved in each set are marked with filled circles and with a red circle for the critical tyrosine in the GRIP domain, and the leucine is in the equivalent position in Rud3p. In both alignments the tryptophans are shaded orange and cluster downstream of the conserved region. In the case of golgin-245, the tryptophan apparently stabilizes the interaction of the GRIP domain with Golgi membranes (Panic et al., 2003a). (C) A schematic representation of the GRAB domain proteins from metazoans and yeasts, along with a GRIP domain protein. All contain either a GRIP or GRAB domain, and the latter have a downstream GA1 motif. Metazoan GRAB proteins also have an extended, proline-rich, COOH-terminal region, whereas GRAB proteins from yeasts and filamentous fungi have an upstream region of sequence conservation (GA2, blue). (D) Fluorescent micrographs of rud3Δ cells expressing GFP-tagged wild-type Rud3p or the mutant L410A as in Fig. 1 A.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172552&req=5

fig2: Rud3p is a member of a family of coiled-coil proteins with a conserved COOH-terminal domain. (A) Schematic representation of S. cerevisiae Rud3p and its relatives from the indicated species. At, A. thaliana; Ce, C. elegans; Dm, D. melanogaster; Hs, Homo sapiens; Sc, S. cerevisiae; Tb, Trypanosoma brucei. (B) Alignment of the COOH-terminal regions of the GRAB domain proteins with those of GRIP domain proteins golgin-245 and golgin-97, and the structure of the GRIP domain of human golgin-245. The two sets of sequences were independently aligned with CLUSTAL W and shaded where more than half the residues are related (gray) or identical (black). Hydrophobic residues conserved in each set are marked with filled circles and with a red circle for the critical tyrosine in the GRIP domain, and the leucine is in the equivalent position in Rud3p. In both alignments the tryptophans are shaded orange and cluster downstream of the conserved region. In the case of golgin-245, the tryptophan apparently stabilizes the interaction of the GRIP domain with Golgi membranes (Panic et al., 2003a). (C) A schematic representation of the GRAB domain proteins from metazoans and yeasts, along with a GRIP domain protein. All contain either a GRIP or GRAB domain, and the latter have a downstream GA1 motif. Metazoan GRAB proteins also have an extended, proline-rich, COOH-terminal region, whereas GRAB proteins from yeasts and filamentous fungi have an upstream region of sequence conservation (GA2, blue). (D) Fluorescent micrographs of rud3Δ cells expressing GFP-tagged wild-type Rud3p or the mutant L410A as in Fig. 1 A.
Mentions: The majority of Rud3p is predicted to form a coiled-coil (Fig. 1 B), but the last 80 residues that mediate targeting lie outside this region. When this COOH-terminal region was used to search the GenBank database using the iterative program PSI-BLAST, it revealed that a large coiled-coil protein with a COOH-terminal domain related to the COOH-terminal 80 residues of Rud3p is present in the genomes of most eukaryotes including mammals, Drosophila melanogaster, Caenorhabditis elegans, plants, fungi, and Dictyostelium discoideum (Fig. 2, A and B). In all but one of these species there is a single member of the family present, the exception being Arabidopsis thaliana, which has two such proteins that are very closely related. The only one of these proteins from other species to have been previously characterized is that from humans, which is called GMAP-210, CEV14, Trip11, or Trip230. This protein was independently identified by either screening an expression library with a human auto-antiserum (GMAP-210, p210; Rios et al., 1994) or by yeast two-hybrid screens with thyroid receptor (Trip11; Lee et al., 1995) and retinoblastoma protein (Trip230; Chang et al., 1997), or as a gene found fused to that of the platelet-derived growth factor receptor by an oncogenic chromosomal translocation (CEV14; Abe et al., 1997). Immunoelectron microscopy has shown that GMAP-210 is localized to the cis side of the Golgi apparatus (Rios et al., 1994).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, England, UK.

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.

Show MeSH
Related in: MedlinePlus