Characterization of the 5′-flanking region of the rat protein kinase C γ gene

Kuang Hua Chen, Steven Widen, Samuel H. Wilson, Kuo Ping Huang

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The 5′-flanking region of protein kinase C (PKC) γ gene was identified from a rat liver genomic library in a bacteriophage λ Charon 4A. A 3.6-kilobase (kb) genomic fragment containing the 5′-flanking region, first exon, and first intron was isolated and sequenced. The transcriptional initiation site, identified by S1 mapping and primer extension, was located 243 base pairs upstream from the translational initiation site. Promoter activity of a DNA segment spanning the 5′-flanking region was demonstrated by both in vitro transcription using HeLa cell nuclear extracts and chloramphenicol acetyltransferase assay by transfection of 293 cells with a PKC γ-CAT fusion construct. Chloramphenicol acetyltransferase assay revealed that a fragment of about 0.16 kb from the transcriptional initiation site was sufficient for promoter activity in these cells, and the construct containing up to 1.6 kb from the cap site was expressed at a similar level. This promoter-active fragment contains several regions similar to defined transcriptional elements in other mammalian promoters, such as those for stimulatory protein 1 (Sp1, activator proteins 1 and 2 (AP1 AP2), c-myc, cAMP regulatory element-binding protein (CREB), and enhancer core (EnhC). Investigation of the genomic structure of PKC 7 gene may lead to the identification of cis-elements controlling tissue-specific and developmental stage-specific expression of PKC γ.

Original languageEnglish (US)
Pages (from-to)19961-19965
Number of pages5
JournalJournal of Biological Chemistry
Volume265
Issue number32
StatePublished - Nov 15 1990
Externally publishedYes

Fingerprint

5' Flanking Region
Protein Kinase C
Rats
Genes
Chloramphenicol O-Acetyltransferase
Assays
Pluto
Bacteriophages
Genomic Library
Transcription Factor AP-1
Transcription
Cell Extracts
HeLa Cells
Base Pairing
Liver
Introns
Transfection
Exons
Carrier Proteins
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of the 5′-flanking region of the rat protein kinase C γ gene. / Chen, Kuang Hua; Widen, Steven; Wilson, Samuel H.; Huang, Kuo Ping.

In: Journal of Biological Chemistry, Vol. 265, No. 32, 15.11.1990, p. 19961-19965.

Research output: Contribution to journalArticle

Chen, Kuang Hua ; Widen, Steven ; Wilson, Samuel H. ; Huang, Kuo Ping. / Characterization of the 5′-flanking region of the rat protein kinase C γ gene. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 32. pp. 19961-19965.
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