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Originally published In Press as doi:10.1074/jbc.M110454200 on December 12, 2001

J. Biol. Chem., Vol. 277, Issue 11, 8866-8876, March 15, 2002
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Transcription Factors NF-Y and Sp1 Are Important Determinants of the Promoter Activity of the Bovine and Human Neuronal Nicotinic Receptor beta 4 Subunit Genes*

Luis M. ValorDagger §, Antonio Campos-CaroDagger §||, Carmen Carrasco-SerranoDagger §**, José A. OrtizDagger §Dagger Dagger , Juan J. Ballesta§§§, and Manuel CriadoDagger §¶¶

From the Departments of Dagger  Biochemistry and Molecular Biology and §§ Pharmacology, and § Instituto de Neurociencias, Universidad Miguel Hernández-C.S.I.C., 03550-San Juan, Alicante, Spain

The beta 4 subunit is a component of the neuronal nicotinic acetylcholine receptors which control catecholamine secretion in bovine adrenomedullary chromaffin cells. The promoter of the gene coding for this subunit was characterized. A proximal region (from -99 to -64) was responsible for the transcriptional activity observed in chromaffin, C2C12, and COS cells. Within this region two cis-acting elements that bind transcription factors Sp1 and NF-Y were identified. Mutagenesis of the two elements indicated that they cooperate for the basal transcription activity of the promoter. The human beta 4 promoter, that was also characterized, shared structural and functional homologies with the bovine promoter. Thus, two adjacent binding elements for Sp1 and NF-Y were detected. Whereas the Sp1 site was an important determinant of the promoter activity, the NF-Y site may have cell-specific effects. Given that these promoters showed no structural or functional homology with the previously characterized rat beta 4 subunit promoter (Bigger, C. B., Casanova, E. A., and Gardner, P. D. (1996) J. Biol. Chem. 271, 32842-32848) except for the involvement of an Sp1 binding element, we propose that constitutive expression of the beta 4 subunit gene in these three close species may be controlled by the general transcription factor Sp1. Nevertheless, other components could determine species-specific beta 4 subunit expression.

* This work was supported in part by Ministry of Education (DGICYT) Grants PB95-0690 and PM98-0104 of Spain, and the Generalitat Valenciana Grant GV-D-VS-20-158-96.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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF453876 and AF453877.

Predoctoral fellow from Generalitat Valenciana.

|| Recipient of a CSIC-Bancaja postdoctoral fellowship.

** Predoctoral fellow from Generalitat Valenciana.

Dagger Dagger Postdoctoral fellow from the Ministry of Education of Spain.

¶¶ To whom correspondence should be addressed. Tel.: 34- 965919479; Fax: 34-965919484; E-mail: Manuel.Criado@umh.es.

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


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