Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region

Jian Wu, Michael J Matunis, Doris Kraemer, Günter Blobel, Elias Coutavas

Research output: Contribution to journalArticle

Abstract

The Ras-related nuclear protein, Ran, has been implicated in nuclear transport. By screening a HeLa cell λ expression library with Ran-GTP and sequencing overlapping cDNA clones, we have obtained the derived primary structure of a protein with a calculated molecular mass of 358 kDa. Using antibodies raised against an expressed segment of this protein, we obtained punctate nuclear surface staining by immunofluorescence microscopy that is characteristic for nucleoporins. Electron microscopy of immunogold-decorated rat liver nuclear envelopes sublocalized the 358-kDa protein at (or near) the tip of the cytoplasmic fibers of the nuclear pore complex (NPC). In agreement with current convention, this protein was therefore termed Nup358 (for nucleoporin of 358 kDa). Nup358 contains a leucine-rich region, four potential Ran binding sites (i.e. Ran binding protein 1 homologous domains) flanked by nucleoporin-characteristic FXFG or FG repeats, eight zinc finger motifs, and a C-terminal cyclophilin A homologous domain. Consistent with the location of Nup358 at the cytoplasmic fibers of the NPC, we found decoration with Ran-gold at only the cytoplasmic side of the NPC. Thus, Nup358 is the first nucleoporin shown to contain binding sites for two of three soluble nuclear transport factors so far isolated, namely karyopherin and Ran-GTP.

Original languageEnglish (US)
Pages (from-to)14209-14213
Number of pages5
JournalJournal of Biological Chemistry
Volume270
Issue number23
StatePublished - Jun 9 1995
Externally publishedYes

Fingerprint

Cyclophilin A
Nuclear Pore Complex Proteins
Zinc Fingers
Guanosine Triphosphate
Leucine
Zinc
Binding Sites
Peptides
Nuclear Pore
Cell Nucleus Active Transport
Proteins
ran GTP-Binding Protein
Karyopherins
Fibers
Nuclear Envelope
Molecular mass
HeLa Cells
Fluorescence Microscopy
Gold
Liver

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region. / Wu, Jian; Matunis, Michael J; Kraemer, Doris; Blobel, Günter; Coutavas, Elias.

In: Journal of Biological Chemistry, Vol. 270, No. 23, 09.06.1995, p. 14209-14213.

Research output: Contribution to journalArticle

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T1 - Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region

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