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PMID: 9230068 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Xlrbpa, a double-stranded RNA-binding protein associated with ribosomes and heterogeneous nuclear RNPs.

The Journal of cell biology ·Vol. 138 ·No. 2 ·1997-07-28 ·Pages 239-53

Eckmann CR, Jantsch MF

Abstract

We have cloned and characterized Xlrbpa, a double-stranded RNA-binding protein from Xenopus laevis. Xlrbpa is a protein of 33 kD and contains three tandemly arranged, double-stranded RNA-binding domains (dsRBDs) that bind exclusively to double-stranded RNA in vitro, but fail to bind either single-stranded RNA or DNA. Sequence data and the overall organization of the protein suggest that Xlrbpa is the Xenopus homologue of human TAR-RNA binding protein (TRBP), a protein isolated by its ability to bind to human immunodeficiency virus (HIV) TAR-RNA. In transfection assays, TRBP has also been shown to inhibit the interferon-induced protein kinase PKR possibly by direct physical interaction. To determine the function of Xlrbpa and its human homologue we studied the expression and intracellular distribution of the two proteins. Xlrbpa is ubiquitously expressed with marked quantitative differences amongst all tissues. Xlrbpa and human TRBP can be detected in the cytoplasm and nucleus by immunofluorescence staining and Western blotting. Sedimentation gradient analyses and immunoprecipitation experiments suggest an association of cytoplasmic Xlrbpa with ribosomes. In contrast, a control construct containing two dsRBDs fails to associate with ribosomes in microinjected Xenopus oocytes. Nuclear staining of Xenopus lampbrush chromosome preparations showed the association of the protein with nucleoli, again indicating an association of the protein with ribosomal RNAs. Additionally, Xlrbpa could be located on lampbrush chromosomes and in snurposomes. Immunoprecipitations of nuclear extracts demonstrated the presence of the protein in heterogeneous nuclear (hn) RNP particles, but not in small nuclear RNPs, explaining the chromosomal localization of the protein. It thus appears that Xlrbpa is a general double-stranded RNA-binding protein which is associated with the majority of cellular RNAs, ribosomal RNAs, and hnRNAs either alone or as part of an hnRNP complex.

MeSH Terms
Amino Acid Sequence Animals Cell Nucleus/chemistry Cloning, Molecular Cytoplasm/chemistry HeLa Cells Heterogeneous-Nuclear Ribonucleoproteins Humans Molecular Sequence Data Molecular Weight Oocytes/chemistry Organ Specificity RNA, Double-Stranded/metabolism RNA-Binding Proteins/analysis,chemistry,genetics,metabolism Recombinant Fusion Proteins/analysis Ribonucleoproteins/metabolism Ribosomal Proteins/metabolism Ribosomes/metabolism Sequence Analysis, DNA Sequence Homology, Amino Acid Xenopus Proteins Xenopus laevis/genetics
Chemicals
Heterogeneous-Nuclear Ribonucleoproteins RNA, Double-Stranded RNA-Binding Proteins Recombinant Fusion Proteins Ribonucleoproteins Ribosomal Proteins Xenopus Proteins Xlrbpa protein, Xenopus trans-activation responsive RNA-binding protein
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Eckmann C R
Department of Cytology and Genetics, Institute of Botany, University of Vienna, A-1030 Vienna, Austria.
Jantsch M F
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.138.2.239
Published
1997-07-28
Pages
239-53
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2138193
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.138.2.239 PMC Full Text View journal details
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