The human chorionic somatomammotropin gene enhancer is composed of multiple DNA elements that are homologous to several SV40 enhansons

Shi Wen Jiang, Norman L. Eberhardt

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Previous studies indicate that a human chorionic somatomammotropin (hCS) gene enhancer (CSEn) associated with the growth hormone (hGH) gene locus is involved in directing cell-specific expression of the hCS genes in placenta. In the current studies, we report a detailed structural analysis of this enhancer. CSEn stimulated transcription of a variety of promoters, including the hCS, human growth hormone, thymidine kinase, and Rous sarcoma virus promoters, in human choriocarcinoma cell lines (BeWo and JEG-3) but not HeLa cells or rat somatolactotrophes (GC). Maximal enhancer activity was confined to a 242-base pair DNA segment. Of several CSEn subfragments, only the En 57/242 subfragment retained activity (33.5% wild-type). The CSEn DNA sequence contained direct and inverted repeat motifs and sequences related to the SV40 enhansons, GT-IIC, GT-I, and SphI/SphII. DNase I footprint analysis revealed that most of these sites were protected by nuclear proteins derived from BeWo, JEG-3, HeLa, and GC cells. Site-specific block mutation of the GT-IIC- related and inverted repeat motifs virtually abolished enhancer activity, and mutation of all but the GT-I-related motif resulted in significant loss (30- 60%) of activity. These data demonstrate that the CS enhancer is comprised of multiple elements related to SV40 enhansons that interact cooperatively to generate enhancer function.

Original languageEnglish (US)
Pages (from-to)10384-10392
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number14
StatePublished - Apr 8 1994

Fingerprint

Placental Lactogen
Genes
DNA
HeLa Cells
Inverted Repeat Sequences
Rous sarcoma virus
Choriocarcinoma
Mutation
Thymidine Kinase
Human Growth Hormone
Nucleic Acid Repetitive Sequences
Deoxyribonuclease I
DNA sequences
Transcription
Nuclear Proteins
Viruses
Structural analysis
Base Pairing
Placenta
Growth Hormone

ASJC Scopus subject areas

  • Biochemistry

Cite this

The human chorionic somatomammotropin gene enhancer is composed of multiple DNA elements that are homologous to several SV40 enhansons. / Jiang, Shi Wen; Eberhardt, Norman L.

In: Journal of Biological Chemistry, Vol. 269, No. 14, 08.04.1994, p. 10384-10392.

Research output: Contribution to journalArticle

@article{632f9014c8754f14b73358c626ff05c7,
title = "The human chorionic somatomammotropin gene enhancer is composed of multiple DNA elements that are homologous to several SV40 enhansons",
abstract = "Previous studies indicate that a human chorionic somatomammotropin (hCS) gene enhancer (CSEn) associated with the growth hormone (hGH) gene locus is involved in directing cell-specific expression of the hCS genes in placenta. In the current studies, we report a detailed structural analysis of this enhancer. CSEn stimulated transcription of a variety of promoters, including the hCS, human growth hormone, thymidine kinase, and Rous sarcoma virus promoters, in human choriocarcinoma cell lines (BeWo and JEG-3) but not HeLa cells or rat somatolactotrophes (GC). Maximal enhancer activity was confined to a 242-base pair DNA segment. Of several CSEn subfragments, only the En 57/242 subfragment retained activity (33.5{\%} wild-type). The CSEn DNA sequence contained direct and inverted repeat motifs and sequences related to the SV40 enhansons, GT-IIC, GT-I, and SphI/SphII. DNase I footprint analysis revealed that most of these sites were protected by nuclear proteins derived from BeWo, JEG-3, HeLa, and GC cells. Site-specific block mutation of the GT-IIC- related and inverted repeat motifs virtually abolished enhancer activity, and mutation of all but the GT-I-related motif resulted in significant loss (30- 60{\%}) of activity. These data demonstrate that the CS enhancer is comprised of multiple elements related to SV40 enhansons that interact cooperatively to generate enhancer function.",
author = "Jiang, {Shi Wen} and Eberhardt, {Norman L.}",
year = "1994",
month = "4",
day = "8",
language = "English (US)",
volume = "269",
pages = "10384--10392",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - The human chorionic somatomammotropin gene enhancer is composed of multiple DNA elements that are homologous to several SV40 enhansons

AU - Jiang, Shi Wen

AU - Eberhardt, Norman L.

PY - 1994/4/8

Y1 - 1994/4/8

N2 - Previous studies indicate that a human chorionic somatomammotropin (hCS) gene enhancer (CSEn) associated with the growth hormone (hGH) gene locus is involved in directing cell-specific expression of the hCS genes in placenta. In the current studies, we report a detailed structural analysis of this enhancer. CSEn stimulated transcription of a variety of promoters, including the hCS, human growth hormone, thymidine kinase, and Rous sarcoma virus promoters, in human choriocarcinoma cell lines (BeWo and JEG-3) but not HeLa cells or rat somatolactotrophes (GC). Maximal enhancer activity was confined to a 242-base pair DNA segment. Of several CSEn subfragments, only the En 57/242 subfragment retained activity (33.5% wild-type). The CSEn DNA sequence contained direct and inverted repeat motifs and sequences related to the SV40 enhansons, GT-IIC, GT-I, and SphI/SphII. DNase I footprint analysis revealed that most of these sites were protected by nuclear proteins derived from BeWo, JEG-3, HeLa, and GC cells. Site-specific block mutation of the GT-IIC- related and inverted repeat motifs virtually abolished enhancer activity, and mutation of all but the GT-I-related motif resulted in significant loss (30- 60%) of activity. These data demonstrate that the CS enhancer is comprised of multiple elements related to SV40 enhansons that interact cooperatively to generate enhancer function.

AB - Previous studies indicate that a human chorionic somatomammotropin (hCS) gene enhancer (CSEn) associated with the growth hormone (hGH) gene locus is involved in directing cell-specific expression of the hCS genes in placenta. In the current studies, we report a detailed structural analysis of this enhancer. CSEn stimulated transcription of a variety of promoters, including the hCS, human growth hormone, thymidine kinase, and Rous sarcoma virus promoters, in human choriocarcinoma cell lines (BeWo and JEG-3) but not HeLa cells or rat somatolactotrophes (GC). Maximal enhancer activity was confined to a 242-base pair DNA segment. Of several CSEn subfragments, only the En 57/242 subfragment retained activity (33.5% wild-type). The CSEn DNA sequence contained direct and inverted repeat motifs and sequences related to the SV40 enhansons, GT-IIC, GT-I, and SphI/SphII. DNase I footprint analysis revealed that most of these sites were protected by nuclear proteins derived from BeWo, JEG-3, HeLa, and GC cells. Site-specific block mutation of the GT-IIC- related and inverted repeat motifs virtually abolished enhancer activity, and mutation of all but the GT-I-related motif resulted in significant loss (30- 60%) of activity. These data demonstrate that the CS enhancer is comprised of multiple elements related to SV40 enhansons that interact cooperatively to generate enhancer function.

UR - http://www.scopus.com/inward/record.url?scp=0028276955&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028276955&partnerID=8YFLogxK

M3 - Article

C2 - 8144621

AN - SCOPUS:0028276955

VL - 269

SP - 10384

EP - 10392

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -