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PMID: 10085285 Published · ppublish English Journal Article

Proteins connecting the nuclear pore complex with the nuclear interior.

The Journal of cell biology ·Vol. 144 ·No. 5 ·1999-03-08 ·Pages 839-55

Strambio-de-Castillia C, Blobel G, Rout MP

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.

MeSH Terms
Amino Acid Sequence Biological Transport Cell Nucleus/metabolism Cloning, Molecular Fungal Proteins/chemistry,genetics,metabolism Gene Deletion Green Fluorescent Proteins Luminescent Proteins/metabolism Microscopy, Electron Molecular Sequence Data Nuclear Envelope/metabolism Nuclear Pore Complex Proteins Nuclear Proteins/chemistry,genetics,metabolism RNA-Binding Proteins Recombinant Fusion Proteins/genetics,metabolism Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid
Chemicals
Fungal Proteins Luminescent Proteins MLP1 protein, S cerevisiae MLP2 protein, S cerevisiae Nuclear Pore Complex Proteins Nuclear Proteins RNA-Binding Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Strambio-de-Castillia C
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York 10021, USA.
Blobel G
Rout M P
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1999-03-08
Pages
839-55
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2148185
Subset
IM
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