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PMID: 11684705 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Novel vertebrate nucleoporins Nup133 and Nup160 play a role in mRNA export.

The Journal of cell biology ·Vol. 155 ·No. 3 ·2001-10-29 ·Pages 339-54

Vasu S, Shah S, Orjalo A, Park M, Fischer WH, Forbes DJ

Abstract

RNA undergoing nuclear export first encounters the basket of the nuclear pore. Two basket proteins, Nup98 and Nup153, are essential for mRNA export, but their molecular partners within the pore are largely unknown. Because the mechanism of RNA export will be in question as long as significant vertebrate pore proteins remain undiscovered, we set out to find their partners. Fragments of Nup98 and Nup153 were used for pulldown experiments from Xenopus egg extracts, which contain abundant disassembled nuclear pores. Strikingly, Nup98 and Nup153 each bound the same four large proteins. Purification and sequence analysis revealed that two are the known vertebrate nucleoporins, Nup96 and Nup107, whereas two mapped to ORFs of unknown function. The genes encoding the novel proteins were cloned, and antibodies were produced. Immunofluorescence reveals them to be new nucleoporins, designated Nup160 and Nup133, which are accessible on the basket side of the pore. Nucleoporins Nup160, Nup133, Nup107, and Nup96 exist as a complex in Xenopus egg extracts and in assembled pores, now termed the Nup160 complex. Sec13 is prominent in Nup98 and Nup153 pulldowns, and we find it to be a member of the Nup160 complex. We have mapped the sites that are required for binding the Nup160 subcomplex, and have found that in Nup98, the binding site is used to tether Nup98 to the nucleus; in Nup153, the binding site targets Nup153 to the nuclear pore. With transfection and in vivo transport assays, we find that specific Nup160 and Nup133 fragments block poly[A]+ RNA export, but not protein import or export. These results demonstrate that two novel vertebrate nucleoporins, Nup160 and Nup133, not only interact with Nup98 and Nup153, but themselves play a role in mRNA export.

MeSH Terms
Active Transport, Cell Nucleus Amino Acid Sequence Animals Binding Sites Cell Nucleus/metabolism HeLa Cells Humans Mice Minor Histocompatibility Antigens Molecular Sequence Data Nuclear Pore/metabolism Nuclear Pore Complex Proteins/genetics,isolation & purification,metabolism,physiology Nuclear Proteins Peptide Fragments RNA, Messenger/metabolism Rats Sequence Homology, Amino Acid Vertebrates Xenopus Xenopus Proteins
Chemicals
Minor Histocompatibility Antigens NUP107 protein, human NUP133 protein, human NUP153 protein, human NUP160 protein, human Nuclear Pore Complex Proteins Nuclear Proteins Nup107 protein, rat Nup153 protein, mouse Nup153 protein, rat Nup160 protein, Xenopus Peptide Fragments RNA, Messenger Xenopus Proteins nuclear pore complex protein 96 nuclear pore complex protein 98
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Vasu S
Section of Cell and Developmental Biology, Division of Biology 0347, University of California at San Diego, La Jolla, CA 92093, USA.
Shah S
Orjalo A
Park M
Fischer W H
Forbes D J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200108007
Published
2001-10-29
Epub
2001-00-29
Pages
339-54
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2150853
Subset
IM
Grants
NIGMS NIH HHS · R01 GM033279 · United States
NIGMS NIH HHS · R01 GM33279 · United States
NCRR NIH HHS · S10 RR 11404-01A1 · United States
Databases
GENBANK
AK001676, AK001754, AW635680, BF048903, BF049549
External Links
PubMed source DOI: 10.1083/jcb.200108007 PMC Full Text View journal details
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