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Title: Cyclic AMP regulation of the human glycoprotein hormone. cap alpha. -subunit gene is mediated by an 18-base-pair element

Abstract

cAMP regulates transcription of the gene encoding the ..cap alpha..-subunit of human chorionic gonadotropin (hCG) in the choriocarcinoma cells (BeWo). To define the sequences required for regulation by cAMP, the authors inserted fragments from the 5' flanking region of the ..cap alpha..-subunit gene into a test vector containing the simian virus 40 early promoter (devoid of its enhancer) linked to the bacterial chloramphenicol acetyltransferase (CAT) gene. Results from transient expression assays in BeWo cells indicated that a 1500-base-pair (bp) fragment conferred cAMP responsiveness on the CAT gene regardless of position or orientation of the insert relative to the viral promoter. A subfragment extending from position -169 to position -100 had the same effect on cAMP-induced expression. Furthermore, the entire stimulatory effect could be achieved with an 18-bp synthetic oligodeoxynucleotide corresponding to a direct repeat between position -146 and -111. In the absence of cAMP, the ..cap alpha..-subunit 5' flanking sequence also enhanced transcription from the simian virus 40 early promoter. They localized this enhancer activity to the same -169/-100 fragment containing the cAMP response element. The 18-bp element alone, however, had no effect on basal expression. Thus, this short DNA sequence serves as a cAMP response element and also functionsmore » independently of other promoter-regulatory elements located in the 5' flanking sequence of the ..cap alpha..-subunit gene.« less

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Case Western Reserve Univ., Cleveland, OH
OSTI Identifier:
6072740
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. U.S.A.; (United States)
Additional Journal Information:
Journal Volume: 84:8
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; AMP; BIOCHEMISTRY; HCG; GENE REGULATION; CARBON 14 COMPOUNDS; GENES; GLUCOPROTEINS; THIN-LAYER CHROMATOGRAPHY; TRANSCRIPTION; VIRUSES; CARBOHYDRATES; CHEMISTRY; CHROMATOGRAPHY; GONADOTROPINS; HORMONES; LABELLED COMPOUNDS; MICROORGANISMS; NUCLEOTIDES; ORGANIC COMPOUNDS; PARASITES; PEPTIDE HORMONES; PITUITARY HORMONES; PROTEINS; SACCHARIDES; SEPARATION PROCESSES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Silver, B J, Bokar, J A, Virgin, J B, Vallen, E A, Milsted, A, and Nilson, J H. Cyclic AMP regulation of the human glycoprotein hormone. cap alpha. -subunit gene is mediated by an 18-base-pair element. United States: N. p., 1987. Web. doi:10.1073/pnas.84.8.2198.
Silver, B J, Bokar, J A, Virgin, J B, Vallen, E A, Milsted, A, & Nilson, J H. Cyclic AMP regulation of the human glycoprotein hormone. cap alpha. -subunit gene is mediated by an 18-base-pair element. United States. https://doi.org/10.1073/pnas.84.8.2198
Silver, B J, Bokar, J A, Virgin, J B, Vallen, E A, Milsted, A, and Nilson, J H. 1987. "Cyclic AMP regulation of the human glycoprotein hormone. cap alpha. -subunit gene is mediated by an 18-base-pair element". United States. https://doi.org/10.1073/pnas.84.8.2198.
@article{osti_6072740,
title = {Cyclic AMP regulation of the human glycoprotein hormone. cap alpha. -subunit gene is mediated by an 18-base-pair element},
author = {Silver, B J and Bokar, J A and Virgin, J B and Vallen, E A and Milsted, A and Nilson, J H},
abstractNote = {cAMP regulates transcription of the gene encoding the ..cap alpha..-subunit of human chorionic gonadotropin (hCG) in the choriocarcinoma cells (BeWo). To define the sequences required for regulation by cAMP, the authors inserted fragments from the 5' flanking region of the ..cap alpha..-subunit gene into a test vector containing the simian virus 40 early promoter (devoid of its enhancer) linked to the bacterial chloramphenicol acetyltransferase (CAT) gene. Results from transient expression assays in BeWo cells indicated that a 1500-base-pair (bp) fragment conferred cAMP responsiveness on the CAT gene regardless of position or orientation of the insert relative to the viral promoter. A subfragment extending from position -169 to position -100 had the same effect on cAMP-induced expression. Furthermore, the entire stimulatory effect could be achieved with an 18-bp synthetic oligodeoxynucleotide corresponding to a direct repeat between position -146 and -111. In the absence of cAMP, the ..cap alpha..-subunit 5' flanking sequence also enhanced transcription from the simian virus 40 early promoter. They localized this enhancer activity to the same -169/-100 fragment containing the cAMP response element. The 18-bp element alone, however, had no effect on basal expression. Thus, this short DNA sequence serves as a cAMP response element and also functions independently of other promoter-regulatory elements located in the 5' flanking sequence of the ..cap alpha..-subunit gene.},
doi = {10.1073/pnas.84.8.2198},
url = {https://www.osti.gov/biblio/6072740}, journal = {Proc. Natl. Acad. Sci. U.S.A.; (United States)},
number = ,
volume = 84:8,
place = {United States},
year = {Wed Apr 01 00:00:00 EST 1987},
month = {Wed Apr 01 00:00:00 EST 1987}
}