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Published online 30 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2411-2420
© 2004 Oxford University Press

The composition of Staufen-containing RNA granules from human cells indicates their role in the regulated transport and translation of messenger RNAs

Patricia Villacé, Rosa M. Marión and Juan Ortín*

Centro Nacional de Biotecnología (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain

*To whom correspondence should be addressed. Tel: +1 34 91 585 4557; Fax: +1 34 91 585 4506; Email: jortin{at}cnb.uam.es
Present address:
Rosa M. Marión, Department of Biochemistry and Biophysics, University of California San Francisco, Room S476, Genentech Hall, 600 16th Street, San Francisco, CA 94143-0448, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Received January 27, 2004; Revised March 16, 2004; Accepted March 30, 2004

hStaufen is the human homolog of dmStaufen, a double-stranded (ds)RNA-binding protein involved in early development of the fly. hStaufen-containing complexes were purified by affinity chromatography from human cells transfected with a TAP-tagged hStaufen gene. These complexes showed a size >10 MDa. Untagged complexes with similar size were identified from differentiated human neuroblasts. The identity of proteins present in purified hStaufen complexes was determined by mass spectrometry and the presence of these proteins and other functionally related ones was verified by western blot. Ribosomes and proteins involved in the control of protein synthesis (PABP1 and FMRP) were present in purified hStaufen complexes, as well as elements of the cytoskeleton (tubulins, tau, actin and internexin), cytoskeleton control proteins (IQGAP1, cdc42 and rac1) and motor proteins (dynein, kinesin and myosin). In addition, proteins normally found in the nucleus, like nucleolin and RNA helicase A, were also found associated with cytosolic hStaufen complexes. The co-localization of these components with hStaufen granules in the dendrites of differentiated neuroblasts, determined by confocal immunofluorescence, validated their association in living cells. These results support the notion that the hStaufen-containing granules are structures essential in the localization and regulated translation of human mRNAs in vivo.


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