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Nucleic Acids Res ; 31(5): 1470-80, 2003 Mar 01.
Article in English | MEDLINE | ID: mdl-12595555

ABSTRACT

The RNA helicases p68 and p72 are highly related members of the DEAD box family of proteins, sharing 90% identity across the conserved core, and have been shown to be involved in both transcription and mRNA processing. We previously showed that these proteins co-localise in the nucleus of interphase cells. In this study we show that p68 and p72 can interact with each other and self-associate in the yeast two-hybrid system. Co-immunoprecipitation experiments confirmed that p68 and p72 can interact in the cell and indicated that these proteins preferentially exist as hetero-dimers. In addition, we show that p68 can interact with NFAR-2, a protein that is also thought to function in mRNA processing. Moreover, gel filtration analysis suggests that p68 and p72 can exist in a variety of complexes in the cell (ranging from approximately 150 to approximately 400 kDa in size), with a subset of p68 molecules being in very large complexes (>2 MDa). The potential to exist in different complexes that may contain p68 and/or p72, together with a range of other factors, would provide the potential for these proteins to interact with different RNA substrates and would be consistent with recent reports implying a wide range of functions for p68/p72.


Subject(s)
Adenosine Triphosphatases/metabolism , Phosphoproteins , Protein Kinases/metabolism , RNA Helicases/metabolism , Adenosine Triphosphatases/chemistry , Adenosine Triphosphatases/genetics , Binding, Competitive , Cell Line , Chromosomal Proteins, Non-Histone/metabolism , DEAD-box RNA Helicases , Dimerization , HeLa Cells , Humans , Microscopy, Fluorescence , Nuclear Factor 90 Proteins , Precipitin Tests , Protein Binding , Protein Kinases/chemistry , Protein Kinases/genetics , RNA Helicases/chemistry , RNA Helicases/genetics , RNA-Binding Proteins/metabolism , Saccharomyces cerevisiae/genetics , Two-Hybrid System Techniques
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