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PMID:8755533
| Citation |
Traglia, HM, O'Connor, JP, Tung, KS, Dallabrida, S, Shen, WC and Hopper, AK (1996) Nucleus-associated pools of Rna1p, the Saccharomyces cerevisiae Ran/TC4 GTPse activating protein involved in nucleus/cytosol transit. Proc. Natl. Acad. Sci. U.S.A. 93:7667-72 |
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| Abstract |
Rna1p is the GTPase activating enzyme for Ran/TC4, a Ras-like GTPase necessary for nuclear/cytosolic exchange. Although most wild-type Rna1p is located in the cytosol, we found that the vast majority of the mutant Rna1-1p and, under appropriate physiological conditions, a small portion of the wild-type Rna1p cofractionate with yeast nuclei. Subnuclear fractionation studies show that most of the Rna1p is tightly associated with nuclear components, and that a portion of the active protein can be solubilized by treatments that fail to solubilize inactive Rna1-1p. To learn the precise nuclear locations of the Rna1 proteins, we studied their subcellular distributions in HeLa cells. By indirect immuno-fluorescence we show that wild-type Rna1p has three subcellular locations. The majority of the protein is distributed throughout the cytosol, but a portion of the protein is nucleus-associated, located at both the cytosolic surface and within the nucleoplasm. Mutant Rna1-1p is found at the outer nuclear surface and in the cytosol. We propose that a small pool of the wild-type Rna1p is located in the nuclear interior, supporting the model that the same components of the Ran/TC4 GTPase cycle exist on both sides of the nuclear membrane. |
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| Keywords |
Base Sequence; Cell Fractionation; Cell Nucleus/metabolism; Cytosol/metabolism; DNA Primers; Fluorescent Antibody Technique, Indirect; GTP-Binding Proteins/analysis; GTP-Binding Proteins/biosynthesis; GTP-Binding Proteins/metabolism; GTPase-Activating Proteins; HeLa Cells; Humans; Models, Biological; Molecular Sequence Data; Mutagenesis, Site-Directed; Nuclear Proteins/metabolism; Open Reading Frames; Polymerase Chain Reaction; Proteins/metabolism; Recombinant Proteins/analysis; Recombinant Proteins/biosynthesis; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/physiology; Saccharomyces cerevisiae Proteins; Sequence Tagged Sites; Transfection; ran GTP-Binding Protein; ras GTPase-Activating Proteins |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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