Purification and Properties of HuD, a Neuronal RNA-binding Protein (*)
- From the (1)Program in Molecular Pharmacology and Therapeutics and the
- (2)Program in Cellular Biochemistry and Biophysics, Memorial Sloan Kettering Cancer Center, New York, New York 10021
- § To whom correspondence should be addressed: Box 20, Laboratory of Molecular Neuro-oncology, Memorial Sloan Kettering Cancer Center, 1275 York Ave., New York, NY 10021. Tel.: 212-639-8701; Fax: 212-794-4332.
Abstract
HuD is a human neuronal specific RNA-binding protein. In this study we have purified HuD and examined its RNA binding properties in detail. HuD binds to mRNAs that contain an AU-rich element with high affinity. In the case of the c-fos AU-rich element, HuD binds to a 27-nucleotide core element comprising AUUUA, AUUUUA, and AUUUUUA motifs. Mutation in any two of these motifs abrogates binding. HuD contains two tandem RNA recognition motifs (RRM), a basic domain, and a third RRM. Deletion analysis has shown that only the first and second RRMs are essential for RNA binding. Thus, these specific RNA binding properties support the idea that the HuD regulates gene expression at the post-transcriptional level.
Footnotes
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↵* This work was supported by National Institutes of Health Grant NS29682 (to H. M. F.) and NCI, National Institutes of Health, Core Grant P30-CA08748. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵1 The abbreviations used are:
- RRM
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RNA recognition motif
- UTR
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untranslated region
- ARE
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AU-rich element.
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↵2W.-J. Ma, S. Chung, and H. M. Furneaux, manuscript in preparation.
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- Received November 14, 1995.
- Revision received February 21, 1996.
- © 1996 by The American Society for Biochemistry and Molecular Biology, Inc.
