Functional Analysis of an Oxygen-Regulated Transcriptional Enhancer Lying 3' to the Mouse Erythropoietin Gene

Erythropoietin, the major hormone controlling red-cell production, is regulated in part through oxygendependent changes in the rate of transcription of its gene. Using transient transfection in HepG2 cells, we have defined a DNA sequence, located 120 base pairs 3' to the poly(A)-addition site o...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1991-12, Vol.88 (23), p.10553-10557
Main Authors: Pugh, C. W., Tan, C. C., Jones, R. W., Ratcliffe, P. J.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Cell Line
Cell lines
CHO Cells
Cloning, Molecular
Cobalt - pharmacology
Cricetinae
Cyanides
Cyanides - pharmacology
Deoxyglucose - pharmacology
Enhancer Elements, Genetic - drug effects
Epics
Erythropoietin - biosynthesis
Erythropoietin - genetics
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression Regulation - drug effects
Genes
Globins - biosynthesis
Globins - genetics
Hep G2 cells
Hypoxia
Kidneys
Male
Mice
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Oxygen - pharmacology
Plasmids
Recombinant Fusion Proteins - biosynthesis
Restriction Mapping
RNA, Messenger - genetics
RNA, Messenger - isolation & purification
Transcription, Genetic - drug effects
Transfection
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Summary:Erythropoietin, the major hormone controlling red-cell production, is regulated in part through oxygendependent changes in the rate of transcription of its gene. Using transient transfection in HepG2 cells, we have defined a DNA sequence, located 120 base pairs 3' to the poly(A)-addition site of the mouse erythropoietin gene, that confers oxygen-regulated expression on a variety of heterologous promoters. The sequence has the typical features of a eukaryotic enhancer. Approximately 70 base pairs are necessary for full activity, but reiteration restores activity to shorter inactive sequences. This enhancer operates in HepG2 and Hep3B cells, but not in Chinese hamster ovary cells or mouse erythroleukemia cells, and responds to cobalt but not to cyanide or 2-deoxyglucose, thus reflecting the physiological control of erythropoietin production accurately.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.88.23.10553