In Drosophila oocytes and neuroblasts, the double-stranded RNA binding protein Staufen assembles into ribonucleoprotein particles, which mediate cytoplasmic mRNA trafficking and translation. Two different mammalian orthologues also appear to reside in distinct RNA-containing particles. To date, relatively little is known about the molecular composition of Staufen-containing ribonucleoprotein complexes. Here, we have used a novel one-step affinity purification protocol to identify components of Staufen 1-containing particles. Whereas the nucleocytoplasmic RNA-binding protein nucleolin is linked to Staufen in an RNA-dependent manner, the association of protein phosphatase 1, the microtubule-dependent motor protein kinesin and several components of the large and small ribosomal subunits with Staufen ribonucleoprotein complexes is RNA-independent. Notably, all these components do not co-purify with a second RNA-binding protein, hnRNPK (heterogeneous ribonucleoprotein K), demonstrating the high specificity of the purification protocol. Furthermore, pull-down and immunoprecipitation experiments suggest a direct interaction between Staufen 1 and the ribosomal protein P0 in vitro as well as in cells. In cell fractionation and sucrose gradient assays, Staufen co-fractionates with intact ribosomes and polysomes, but not with the isolated 40 S ribosomal subunit. Taken together, these findings imply that, in the cytoplasm of mammalian cells, an association with the ribosomal P-stalk protein P0 recruits Staufen 1 into ribosome-containing ribonucleoprotein particles, which also contain kinesin, protein phosphatase 1 and nucleolin.
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Research Article|
November 23 2004
Characterization of Staufen 1 ribonucleoprotein complexes
Cornelia BRENDEL;
Cornelia BRENDEL
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
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Monika REHBEIN;
Monika REHBEIN
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
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Hans-Jürgen KREIENKAMP;
Hans-Jürgen KREIENKAMP
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
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Friedrich BUCK;
Friedrich BUCK
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
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Dietmar RICHTER;
Dietmar RICHTER
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
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Stefan KINDLER
Stefan KINDLER
1
1Institute for Cell Biochemistry and Clinical Neurobiology, University Hospital Hamburg-Eppendorf, D-20246 Hamburg, Germany
1To whom correspondence should be addressed (email kindler@uke.uni-hamburg.de).
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Publisher: Portland Press Ltd
Received:
May 17 2004
Revision Received:
July 21 2004
Accepted:
August 10 2004
Accepted Manuscript online:
August 10 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2004
Biochem J (2004) 384 (2): 239–246.
Article history
Received:
May 17 2004
Revision Received:
July 21 2004
Accepted:
August 10 2004
Accepted Manuscript online:
August 10 2004
Citation
Cornelia BRENDEL, Monika REHBEIN, Hans-Jürgen KREIENKAMP, Friedrich BUCK, Dietmar RICHTER, Stefan KINDLER; Characterization of Staufen 1 ribonucleoprotein complexes. Biochem J 1 December 2004; 384 (2): 239–246. doi: https://doi.org/10.1042/BJ20040812
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