Transcription of the human ferredoxin gene through a single promoter which contains the 3',5'-cyclic adenosine monophosphate-responsive sequence and Sp 1-binding site

Chi Yao Chang, Cory Huang, Ing Cherng Guo, Huei Man Tsai, Du An Wu, Bon Chu Chung

Research output: Contribution to journalArticle

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Abstract

We have investigated the functional elements involved in cAMP-stimulated transcription of the human ferredoxin gene. Unlike the bovine gene, the human gene lacked a second upstream RNA initiation site as demonstrated by sequence analysis of the exon boundary, lack of upstream RNA, and analysis of the promoter. The presence of a single promoter was determined by testing the ability of various gene segments to drive the expression of the chloramphenicol acetyltransferase gene after transfection into a mouse adrenal cell line Y1. Full promoter activity was conferred by a DNA fragment spanning -209 to +55, although the -94 to +55 fragment already provided some promoter activity. Transcription from the -94 to +55 segment was stimulated by 2-fold when 8-bromo-cAMP was added to the cell. Footprinting analyses showed two GC boxes at -50 to -70 and -87 to -108 were protected by proteins from both Y1 and HeLa cells. Competition experiments showed that a protein with a recognition sequence indistinguishable from Sp1 bound to these sites. When connected to a heterologous TATA box, the sequence at -76 to -42, which contained the proximal GC box, was able to confer a high level of basal transcription and cAMP stimulation. This sequence does not show sequence homology with the known cAMP-responsive element. Mutations or deletion of the Sp 1-binding site showed diminished basal transcription and defined the cAMP responsive sequence to be from -76 to -62. Therefore the cAMP-responsive sequence of the human ferredoxin gene was located at -76 to -62, which was adjacent to the Sp 1-binding site.

Original languageEnglish
Pages (from-to)1362-1370
Number of pages9
JournalMolecular Endocrinology
Volume6
Issue number9
DOIs
Publication statusPublished - Jan 1 1992

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Ferredoxins
Cyclic AMP
Binding Sites
Genes
RNA
8-Bromo Cyclic Adenosine Monophosphate
TATA Box
Chloramphenicol O-Acetyltransferase
Sequence Deletion
Sequence Homology
HeLa Cells
Transfection
Sequence Analysis
Exons
Proteins
Cell Line
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Transcription of the human ferredoxin gene through a single promoter which contains the 3',5'-cyclic adenosine monophosphate-responsive sequence and Sp 1-binding site. / Chang, Chi Yao; Huang, Cory; Guo, Ing Cherng; Tsai, Huei Man; Wu, Du An; Chung, Bon Chu.

In: Molecular Endocrinology, Vol. 6, No. 9, 01.01.1992, p. 1362-1370.

Research output: Contribution to journalArticle

Chang, Chi Yao ; Huang, Cory ; Guo, Ing Cherng ; Tsai, Huei Man ; Wu, Du An ; Chung, Bon Chu. / Transcription of the human ferredoxin gene through a single promoter which contains the 3',5'-cyclic adenosine monophosphate-responsive sequence and Sp 1-binding site. In: Molecular Endocrinology. 1992 ; Vol. 6, No. 9. pp. 1362-1370.
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