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J Biol Chem, Vol. 275, Issue 2, 1307-1314, January 14, 2000

An Element in the Region Responsible for Premature Termination of Transcription Mediates Repression of c-myc Gene Expression by Thyroid Hormone in Neuroblastoma Cells*

Germán Pérez-JusteDagger , Susana García-Silva, and Ana Aranda§

From the Instituto de Investigaciones Biomédicas "Alberto Sols", Consejo Superior de Investigaciones Científicas and Universidad Aut-noma de Madrid, 28029 Madrid, Spain

The thyroid hormone (T3) blocks proliferation and induces differentiation of neuroblastoma N2a-beta cells that express the thyroid hormone receptor (TR) beta 1 isoform. c-Myc is required for cell cycle progression, and this study shows that T3-induced neuronal differentiation is preceded by a rapid decrease of c-myc gene expression. A negative T3 responsive element (TRE), arranged as an inverted palindrome spaced by three nucleotides, has been identified within the first exon between nucleotides +237 and +268. The TRE is adjacent to the binding site for the transcriptional repressor CCCTC binding factor and maps precisely within the region of RNA polymerase II pausing and release, suggesting a direct implication of TR on premature termination of transcription. Furthermore, the TRE confers repression by T3 to an heterologous promoter only when inserted downstream of the transcription initiation site. Binding of CCCTC binding factor and TR to their cognate sites in the region of transcriptional attenuation, as well as direct interactions between both factors, could facilitate the formation of a repressor complex and the inhibition of c-myc gene expression. These studies provide insight into mechanisms by which TR mediate transcriptional repression and contribute to the understanding of the important effects of thyroid hormones on growth and differentiation of neuronal cells.

* This work was supported by Grants PM97-0135 from the Direccion General de Enseñanza Superior and 08.1/0032 from the Comunidad de Madrid.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.

Dagger Fellow from the Comunidad de Madrid.

§ To whom correspondence should be addressed: Instituto de Investigaciones Biomédicas, Arturo Duperier 4, 28029 Madrid, Spain. Tel.: 34-91-5854642; Fax: 34-91-5854587; E-mail: aaranda@iib.uam.es.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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