Cell-specific expression of the rat secretogranin II promoter

Letetia C. Jones, Richard N. Day, Steven J. Pittler, Donna L. Valentine, Jonathan G. Scammell

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Secretogranin II (SgII) is a member of the granin family of secretory proteins, which are selectively expressed in neuroendocrine cells. As a first step in understanding the molecular basis for cell typespecific expression of SgII, we isolated a 12-kb done from a rat genomic library that contained the entire rat SgII coding region, the transcription initiation site, and approximately 3 kb of 5'-flanking region. Within 75 bp of the transcription start site (+1) we located a TATA box and a consensus cAMP responsive element. Within the 5'-flanking region, a number of potential cis-acting elements were identified, including 2 Pit-1 binding sites, 15 E box motifs, and near perfect matches for AP-1 and AP-2 sites. To demonstrate cell type- specific expression the rat SgII gene, a plasmid containing 2.6 kb of the 5'- flanking region of the SgII gene fused to the luciferase reporter gene (p2774Luc) was transfected into rat pheochromocytoma PC-12 cells, rat pituitary GH4C1 (GH) cells, human BE(2)-M17 (M17) neuroblastoma cells, and mouse fibroblast NIH/3T3 cells. The promoter activity was 6- to 36-fold higher in neuroendocrine cells than in NIH/3T3 cells. Progressive deletions in the 5'-flanking region to 61 bp upstream of the start site (p223Luc) had no effect on promoter activity in PC-12 cells. On the other hand, a 5'- deletion in the SgII promoter to -1032 increased promoter activity 3.8-fold in GH cells. This level of expression was maintained when the SgII promoter was further truncated to -189, whereas truncation to -61 resulted in a 2.6- fold reduction in promoter activity. These results suggest that the sequence between -61 and +162 bp is sufficient for SgII promoter activity in PC-12 cells. However, other elements in the 5'-flanking region contribute to both positive and negative regulation of the rat SgII gene in GH cells.

Original languageEnglish (US)
Pages (from-to)3815-3822
Number of pages8
JournalEndocrinology
Volume137
Issue number9
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology

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    Jones, L. C., Day, R. N., Pittler, S. J., Valentine, D. L., & Scammell, J. G. (1996). Cell-specific expression of the rat secretogranin II promoter. Endocrinology, 137(9), 3815-3822. https://doi.org/10.1210/endo.137.9.8756552