Insulin stimulates rat calmodulin I gene transcription through activation of Sp1

S. S. Solomon, M. R. Palazzolo, T. Takahashi, Rajendra Raghow

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Abstract

We have shown previously that insulin positively regulates transcription of the rat calmodulin (CAM) I gene. This activation occurs concomitantly with the activation of the low-Km adenosine 3':5'-cyclic phosphate phosphodiesterase (PDE), which appears to be coregulated with CaM. Rat hepatoma H-411E cells were transfected with plasmids containing various lengths of the putative CaM promoter coupled to a luciferase reporter and were challenged with insulin. We demonstrate that insulin-stimulated transcription of CaM I gene is mediated by a 392-bp 5'-flanking region of the CaM I gene, encompassing 185 bp downstream and 207 bp upstream of the start site of transcription. The CaM I promoter contains three potential Spl sites, located at - 114 through - 109 [(3), +], -77 through -72 [(2), -] and at +53 through +58 [(1), +]. The gel mobility shift assays demonstrated that nuclear protein(s) associate with all three Sp1 sites. We present data demonstrating the relative importance of the three Spl sites for the insulin effect: prCaM I 1835, 3.8X. Δ1081; prCaM I 392, 5.3x, Δ1055; prCaM I 180, 3.7x, Δ462; prCaM I 237, 1.6x, Δ478; prCaM I 139, 2.6x, Δ182; prCaM I 130, 2.1x, Δ194; and prCaM I 1463, negligible activity. In summary, the maximal insulin stimulation of CaM gene expression is seen when the promoter region contains at least two Sp1 sites.

Original languageEnglish (US)
Pages (from-to)470-477
Number of pages8
JournalProceedings of the Association of American Physicians
Volume109
Issue number5
StatePublished - Sep 22 1997
Externally publishedYes

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Calmodulin
Transcriptional Activation
Insulin
Genes
5' Flanking Region
Transcription Initiation Site
Phosphoric Diester Hydrolases
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Luciferases
Genetic Promoter Regions
Adenosine
Hepatocellular Carcinoma
Plasmids
Gels
Phosphates
Gene Expression

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Insulin stimulates rat calmodulin I gene transcription through activation of Sp1. / Solomon, S. S.; Palazzolo, M. R.; Takahashi, T.; Raghow, Rajendra.

In: Proceedings of the Association of American Physicians, Vol. 109, No. 5, 22.09.1997, p. 470-477.

Research output: Contribution to journalArticle

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