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PMID: 10022908 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A human sequence homologue of Staufen is an RNA-binding protein that is associated with polysomes and localizes to the rough endoplasmic reticulum.

Molecular and cellular biology ·Vol. 19 ·No. 3 ·1999-03-00 ·Pages 2212-9

Marión RM, Fortes P, Beloso A, Dotti C, Ortín J

Abstract

In the course of a two-hybrid screen with the NS1 protein of influenza virus, a human clone capable of coding for a protein with high homology to the Staufen protein from Drosophila melanogaster (dmStaufen) was identified. With these sequences used as a probe, cDNAs were isolated from a lambda cDNA library. The encoded protein (hStaufen-like) contained four double-stranded RNA (dsRNA)-binding domains with 55% similarity and 38% identity to those of dmStaufen, including identity at all residues involved in RNA binding. A recombinant protein containing all dsRNA-binding domains was expressed in Escherichia coli as a His-tagged polypeptide. It showed dsRNA binding activity in vitro, with an apparent Kd of 10(-9) M. Using a specific antibody, we detected in human cells a major form of the hStaufen-like protein with an apparent molecular mass of 60 to 65 kDa. The intracellular localization of hStaufen-like protein was investigated by immunofluorescence using a series of markers for the cell compartments. Colocalization was observed with the rough endoplasmic reticulum but not with endosomes, cytoskeleton, or Golgi apparatus. Furthermore, sedimentation analyses indicated that hStaufen-like protein associates with polysomes. These results are discussed in relation to the possible functions of the protein.

MeSH Terms
Amino Acid Sequence Animals COS Cells Cell Line, Transformed Cytoskeletal Proteins Drosophila Proteins Drosophila melanogaster Endoplasmic Reticulum, Rough/metabolism Gene Expression HeLa Cells Humans Molecular Sequence Data Polyribosomes/metabolism RNA-Binding Proteins/genetics,metabolism Viral Nonstructural Proteins/genetics,metabolism
Chemicals
Cytoskeletal Proteins Drosophila Proteins INS1 protein, influenza virus RNA-Binding Proteins STAU1 protein, human Viral Nonstructural Proteins staufen protein, mammalian stau protein, Drosophila
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Marión R M
Centro Nacional de Biotecnología (CSIC), Cantoblanco, 28049 Madrid, Spain.
Fortes P
Beloso A
Dotti C
Ortín J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.3.2212
Published
1999-03-00
Pages
2212-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84014
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.19.3.2212 PMC Full Text View journal details
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