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J. Biol. Chem., Vol. 278, Issue 39, 37249-37255, September 26, 2003

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Transcriptional Regulation by Activation and Repression Elements Located at the 5'-Noncoding Region of the Human 9 Nicotinic Receptor Subunit Gene^*

Luis M. Valor , Mar Castillo , José A. Ortiz ¶ and Manuel Criado ||

From the Department of Biochemistry and Molecular Biology and the Instituto de Neurociencias, Universidad Miguel Hernández-Consejo Superior de Investigaciones Científicas, 03550 San Juan, Alicante, Spain

The 9 subunit is a component of the neuronal nicotinic acetylcholine receptor gene superfamily that is expressed in very restricted locations. The promoter of the human gene has been analyzed in the human neuroblastoma SH-SY5Y, where 9 subunit expression was detected, and in C2C12 cells that do not express 9. A proximal promoter region (from –322 to +113) showed maximal transcriptional activity in SH-SY5Y cells, whereas its activity in C1C12 cells was much lower. Two elements unusually located at the 5'-noncoding region exhibited opposite roles. A negative element located between +15 and +48 appears to be cell-specific because it was effective in C2C12 but not in SH-SY5Y cells, where it was counterbalanced by the presence of the promoter region 5' to the initiation site. An activating element located between +66 and +79 and formed by two adjacent Sox boxes increased the activity of the 9 promoter about 4-fold and was even able to activate other promoters. This element interacts with Sox proteins, probably through a cooperative mechanism in which the two Sox boxes are necessary. We propose that the Sox complex provides an initial scaffold that facilitates the recruiting of the transcriptional machinery responsible for 9 subunit expression.

Received for publication, July 2, 2003 , and in revised form, July 14, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AJ576315.

^* This work was supported by grants from the Ministries of Education (PM98-0104) and Science and Technology (BMC2002-00972) of Spain and from the Generalitat Valenciana (CTIDIB/2002/138). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of predoctoral fellowships from the Generalitat Valenciana and Consejo Superior de Investigaciones Científicas-Bancaja.

Recipient of a fellowship from the Ministry of Science and Technology (MCyT) of Spain.

^¶ Supported by a grant from the MCyT of Spain ("Ramón y Cajal" program).

^|| To whom correspondence should be addressed. Tel.: 34-965919479; Fax: 34-965919484; E-mail: Manuel.Criado{at}umh.es.

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