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PMID: 8557738 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The GLFG repetitive region of the nucleoporin Nup116p interacts with Kap95p, an essential yeast nuclear import factor.

The Journal of cell biology ·Vol. 131 ·No. 6 Pt 2 ·1995-12-00 ·Pages 1699-713

Iovine MK, Watkins JL, Wente SR

Abstract

Nup116p is a member of a family of five yeast nuclear pore complex (NPC) proteins that share an amino terminal region of repetitive tetrapeptide "GLFG" motifs. Previous experiments characterized the unique morphological perturbations that occur in a nup116 null mutant: temperature-sensitive formation of nuclear envelope seals over the cytoplasmic face of the NPC (Wente, S. R., and G. Blobel. 1993. J. Cell Biol. 123:275-284). Three approaches have been taken to dissect the structural basis for Nup116p's role in NPC function. First, deletion mutagenesis analysis of NUP116 revealed that the GLFG region was required for NPC function. This was not true for the other four yeast GLFG family members (Nup49p, Nup57p, Nup100p, and Nup145p). Moreover, deletion of either half of Nup116p's GLFG repeats or replacement of Nup116p's GLFG region with either Nup100p's GLFG region or Nsp1p's FXFG repetitive region abolishes the function of Nup116p. At a semipermissive growth temperature, the cells lacking Nup116p's GLFG region displayed a diminished capacity for nuclear import. Second, overexpression of Nup116p's GLFG region severely inhibited cell growth, rapidly blocked polyadenylated-RNA export, and fragmented the nucleolus. Although it inhibited nuclear export, the overexpressed GLFG region appeared predominantly localized in the cytoplasm and NPC/nuclear envelope structure was not perturbed in thin section electron micrographs. Finally, using biochemical and two-hybrid analysis, an interaction was characterized between Nup116p's GLFG region and Kap95p, an essential yeast homologue of the vertebrate nuclear import factor p97/Imp90/karopherin beta. These data show that Nup116p's GLFG region has an essential role in mediating nuclear transport.

MeSH Terms
Amino Acid Sequence Base Sequence Biological Transport/genetics Cell Division/physiology Cell Nucleus/genetics,metabolism Cell Survival/genetics Fungal Proteins/genetics,metabolism Galactose/pharmacology Gene Deletion Gene Expression/physiology Membrane Proteins/genetics,metabolism Microscopy, Electron Molecular Sequence Data Nuclear Pore Complex Proteins Nuclear Proteins/genetics,metabolism Phenotype RNA, Messenger/metabolism Raffinose/pharmacology Repetitive Sequences, Nucleic Acid/genetics Saccharomyces cerevisiae Proteins Yeasts/cytology,metabolism,ultrastructure
Chemicals
Fungal Proteins Membrane Proteins NUP116 protein, S cerevisiae Nuclear Pore Complex Proteins Nuclear Proteins RNA, Messenger Saccharomyces cerevisiae Proteins Raffinose Galactose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Iovine M K
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Watkins J L
Wente S R
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.131.6.1699
Published
1995-12-00
Pages
1699-713
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120653
Subset
IM
Grants
NIGMS NIH HHS · 2T32GM07067-21 · United States
NIGMS NIH HHS · R01 GM51219-02 · United States
External Links
PubMed source DOI: 10.1083/jcb.131.6.1699 PMC Full Text View journal details
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