Faculty Collaboration Database

RNA association defines a functionally conserved domain in the nuclear pore protein Nup153. J Biol Chem 2001 Nov 30;276(48):45349-57

Date

09/22/2001

Pubmed ID

11567018

DOI

10.1074/jbc.M102592200

Scopus ID

2-s2.0-0035977057 (requires institutional sign-in at Scopus site)   21 Citations

Abstract

Traffic between the nucleus and cytoplasm takes place through a macromolecular structure termed the nuclear pore complex. To understand how the vital process of nucleocytoplasmic transport occurs, the contribution of individual pore proteins must be elucidated. One such protein, the nucleoporin Nup153, is localized to the nuclear basket of the pore complex and has been shown to be a central component of the nuclear transport machinery. Perturbation of Nup153 function was demonstrated previously to block the export of several classes of RNA cargo. Moreover, these studies also showed that Nup153 can stably associate with RNA in vitro. In this study, we have mapped a domain within Nup153, encompassing amino acids 250-400 in human Nup153, that is responsible for RNA association. After cloning this region of Xenopus Nup153, we performed a cross-species analysis. Despite variation in sequence conservation between Drosophila, Xenopus, and human, this domain of Nup153 displayed robust RNA binding activity in each case, indicating that this property is a hallmark feature of Nup153 and pointing toward a subset of amino acid residues that are key to conferring this ability. We have further determined that a recombinant fragment of Nup153 can bind directly to RNA and that this fragment can interact with endogenous RNA targets. Our findings identify a functionally conserved domain in Nup153 and suggest a role for RNA binding in Nup153 function at the nuclear pore.

Author List

Dimaano C, Ball JR, Prunuske AJ, Ullman KS

Author

Amy Jeanette Prunuske PhD Professor in the Medical School Regional Campuses department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Amino Acids
Animals
Blotting, Western
Cell Nucleus
Cloning, Molecular
Conserved Sequence
Drosophila
Evolution, Molecular
Humans
Molecular Sequence Data
Nuclear Pore Complex Proteins
Oocytes
Polymerase Chain Reaction
Protein Binding
Protein Structure, Tertiary
RNA
Recombinant Proteins
Sequence Homology, Amino Acid
Species Specificity
Xenopus
Xenopus Proteins