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PMID:10085285
| Citation |
Strambio-de-Castillia, C, Blobel, G and Rout, MP (1999) Proteins connecting the nuclear pore complex with the nuclear interior. J. Cell Biol. 144:839-55 |
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| Abstract |
While much has been learned in recent years about the movement of soluble transport factors across the nuclear pore complex (NPC), comparatively little is known about intranuclear trafficking. We isolated the previously identified Saccharomyces protein Mlp1p (myosin-like protein) by an assay designed to find nuclear envelope (NE) associated proteins that are not nucleoporins. We localized both Mlp1p and a closely related protein that we termed Mlp2p to filamentous structures stretching from the nucleoplasmic face of the NE into the nucleoplasm, similar to the homologous vertebrate and Drosophila Tpr proteins. Mlp1p can be imported into the nucleus by virtue of a nuclear localization sequence (NLS) within its COOH-terminal domain. Overexpression experiments indicate that Mlp1p can form large structures within the nucleus which exclude chromatin but appear highly permeable to proteins. Remarkably, cells harboring a double deletion of MLP1 and MLP2 were viable, although they showed a slower net rate of active nuclear import and faster passive efflux of a reporter protein. Our data indicate that the Tpr homologues are not merely NPC-associated proteins but that they can be part of NPC-independent, peripheral intranuclear structures. In addition, we suggest that the Tpr filaments could provide chromatin-free conduits or tracks to guide the efficient translocation of macromolecules between the nucleoplasm and the NPC. |
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| Keywords |
Amino Acid Sequence; Biological Transport; Cell Nucleus/metabolism; Cloning, Molecular; Fungal Proteins/chemistry; Fungal Proteins/genetics; Fungal Proteins/metabolism; Gene Deletion; Green Fluorescent Proteins; Luminescent Proteins/metabolism; Microscopy, Electron; Molecular Sequence Data; Nuclear Envelope/metabolism; Nuclear Pore Complex Proteins; Nuclear Proteins/chemistry; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid |
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