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Originally published In Press as doi:10.1074/jbc.M106966200 on November 15, 2001

J. Biol. Chem., Vol. 277, Issue 3, 1967-1973, January 18, 2002
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HuD, a Neuronal-specific RNA-binding Protein, Increases the in Vivo Stability of MYCN RNA*

Chitra F. ManoharDagger , Marc L. ShortDagger §, Anthony NguyenDagger , Nadine N. NguyenDagger , Daniel ChagnovichDagger , Qiwei YangDagger , and Susan L. Cohn||**

From the || Department of Pediatrics and Dagger  The Robert H. Lurie Comprehensive Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611

MYCN amplification and consequent deregulated expression plays a crucial role in determining the clinical behavior of neuroblastoma. Enhanced expression of MYCN confers growth potential to neuroblastoma cells, and a direct link between MYCN expression and the development of neuroblastoma has been demonstrated in transgenic mice studies. Although the molecular pathways underlying the regulation of MYCN have not been fully elucidated, post-transcriptional mechanisms appear to be important. Previously, we reported that an embryonic lethal abnormal vision-like (ELAV) protein binds with high specificity to at least two AU-rich elements within the MYCN 3'-untranslated region. In this study, we characterized the ability of cis-acting elements within the MYCN 3'-untranslated region to destabilize mRNA in cells and examined the functional consequences of its interactions with the ELAV protein HuD. We show that at least 4 cis-acting elements within the MYCN 3'-untranslated region are able to signal the degradation of stable heterologous mRNA. Ectopic overexpression of HuD dramatically inhibits RNA decay mediated by the full-length MYCN 3'-untranslated region and cis-acting destabilizing elements that harbor HuD binding sites in vivo. HuD may contribute to the malignant phenotype of neuroblastoma cells by stabilizing MYCN mRNA, thereby enhancing steady-state levels of expression of this oncogene.

* This work was supported by NCI, National Institutes of Health (NIH) Grant CA74824, the Elise Anderson Neuroblastoma Research Fund, the Neuroblastoma Children's Cancer Society, gifts from Dennis Drescher, and the Robert H. Lurie Comprehensive Cancer Center, NCI, NIH Core Grant 5P30CA60553.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Current address: Applied Biosystems, 850 Lincoln Center Dr., Foster City, CA 94404.

Current address: Skirball Institute, 540 1st Ave., New York, NY 10016.

** To whom correspondence should be addressed: Children's Memorial Hospital, Division of Hematology/Oncology, Box 30, 2300 Children's Plaza, Chicago, IL 60614. Tel.: 773-880-4562; Fax: 773-880-3053; E-mail: scohn@northwestern.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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