Article
- The EMBO Journal (2004) 23, 3346 - 3355
- doi:10.1038/sj.emboj.7600341
Published online: 29 July 2004
Visualization of RNA–protein interactions in living cells: FMRP and IMP1 interact on mRNAs
Oliver Rackhama and Chris M Brown
- Biochemistry Department, University of Otago, Dunedin, New Zealand
Correspondence to:
Chris M Brown, Biochemistry Department, University of Otago, PO Box 56, Dunedin 9001, New Zealand. Tel.: +64 3 47 95 201; Fax: +64 3 47 97 866; E-mail: chris.brown@otago.ac.nz
aPresent address: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK
Received 6 October 2003; Accepted 1 July 2004
Abstract
Protein expression depends significantly on the stability, translation efficiency and localization of mRNA. These qualities are largely dictated by the RNA-binding proteins associated with an mRNA. Here, we report a method to visualize and localize RNA–protein interactions in living mammalian cells. Using this method, we found that the fragile X mental retardation protein (FMRP) isoform 18 and the human zipcode-binding protein 1 ortholog IMP1, an RNA transport factor, were present on common mRNAs. These interactions occurred predominantly in the cytoplasm, in granular structures. In addition, FMRP and IMP1 interacted independently of RNA. Tethering of FMRP to an mRNA caused IMP1 to be recruited to the same mRNA and resulted in granule formation. The intimate association of FMRP and IMP1 suggests a link between mRNA transport and translational repression in mammalian cells.
Keywords:
- fragile X syndrome,
- microscopy, fluorescence,
- RNA-binding proteins,
- RNA transport,
- translation, genetic
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