Characterization of the promoter of the human Gi2 α-subunit gene

Lee S. Weinstein, Inna Kats, Allen M. Spiegel, Anthony D. Carter

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The promoter of the human gene for the α-subunit of Gi2, a GTP-binding signal transduction protein, was analyzed by cloning the 5′ flanking region of the gene upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and measuring the level of CAT expression after transfection in CV-1 green monkey kidney cells. Analysis of multiple 5′ deletion mutants reveals that a minimum of 85 bases upstream of the major transcriptional initiation site are required for full basal promoter activity. Deleting 11 bases to position -74 causes a 4-fold decrease in promoter activity. Another major decrement in activity is seen when a GC box between -46 and -33 is deleted, consistent with this region being a functionally active Sp1 factor-binding site. Primer extension of a CAT-specific primer hybridized to RNA from cells transfected with a Gi2α promoter-CAT construct confirms approximately the same transcriptional start site as the endogenous Gi2α gene. The 3′ deletion mutants that either approach or delete the transcriptional start site have markedly diminished activity.

Original languageEnglish (US)
Pages (from-to)958-964
Number of pages7
JournalMolecular Endocrinology
Volume4
Issue number7
StatePublished - Jul 1990
Externally publishedYes

Fingerprint

Chloramphenicol O-Acetyltransferase
Genes
Bacterial Genes
Cercopithecus aethiops
5' Flanking Region
Guanosine Triphosphate
Transfection
Organism Cloning
Signal Transduction
Binding Sites
RNA
Kidney
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Weinstein, L. S., Kats, I., Spiegel, A. M., & Carter, A. D. (1990). Characterization of the promoter of the human Gi2 α-subunit gene. Molecular Endocrinology, 4(7), 958-964.

Characterization of the promoter of the human Gi2 α-subunit gene. / Weinstein, Lee S.; Kats, Inna; Spiegel, Allen M.; Carter, Anthony D.

In: Molecular Endocrinology, Vol. 4, No. 7, 07.1990, p. 958-964.

Research output: Contribution to journalArticle

Weinstein, LS, Kats, I, Spiegel, AM & Carter, AD 1990, 'Characterization of the promoter of the human Gi2 α-subunit gene', Molecular Endocrinology, vol. 4, no. 7, pp. 958-964.
Weinstein, Lee S. ; Kats, Inna ; Spiegel, Allen M. ; Carter, Anthony D. / Characterization of the promoter of the human Gi2 α-subunit gene. In: Molecular Endocrinology. 1990 ; Vol. 4, No. 7. pp. 958-964.
@article{c35c7792bde2488c986bacea3b4cc988,
title = "Characterization of the promoter of the human Gi2 α-subunit gene",
abstract = "The promoter of the human gene for the α-subunit of Gi2, a GTP-binding signal transduction protein, was analyzed by cloning the 5′ flanking region of the gene upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and measuring the level of CAT expression after transfection in CV-1 green monkey kidney cells. Analysis of multiple 5′ deletion mutants reveals that a minimum of 85 bases upstream of the major transcriptional initiation site are required for full basal promoter activity. Deleting 11 bases to position -74 causes a 4-fold decrease in promoter activity. Another major decrement in activity is seen when a GC box between -46 and -33 is deleted, consistent with this region being a functionally active Sp1 factor-binding site. Primer extension of a CAT-specific primer hybridized to RNA from cells transfected with a Gi2α promoter-CAT construct confirms approximately the same transcriptional start site as the endogenous Gi2α gene. The 3′ deletion mutants that either approach or delete the transcriptional start site have markedly diminished activity.",
author = "Weinstein, {Lee S.} and Inna Kats and Spiegel, {Allen M.} and Carter, {Anthony D.}",
year = "1990",
month = "7",
language = "English (US)",
volume = "4",
pages = "958--964",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "7",

}

TY - JOUR

T1 - Characterization of the promoter of the human Gi2 α-subunit gene

AU - Weinstein, Lee S.

AU - Kats, Inna

AU - Spiegel, Allen M.

AU - Carter, Anthony D.

PY - 1990/7

Y1 - 1990/7

N2 - The promoter of the human gene for the α-subunit of Gi2, a GTP-binding signal transduction protein, was analyzed by cloning the 5′ flanking region of the gene upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and measuring the level of CAT expression after transfection in CV-1 green monkey kidney cells. Analysis of multiple 5′ deletion mutants reveals that a minimum of 85 bases upstream of the major transcriptional initiation site are required for full basal promoter activity. Deleting 11 bases to position -74 causes a 4-fold decrease in promoter activity. Another major decrement in activity is seen when a GC box between -46 and -33 is deleted, consistent with this region being a functionally active Sp1 factor-binding site. Primer extension of a CAT-specific primer hybridized to RNA from cells transfected with a Gi2α promoter-CAT construct confirms approximately the same transcriptional start site as the endogenous Gi2α gene. The 3′ deletion mutants that either approach or delete the transcriptional start site have markedly diminished activity.

AB - The promoter of the human gene for the α-subunit of Gi2, a GTP-binding signal transduction protein, was analyzed by cloning the 5′ flanking region of the gene upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and measuring the level of CAT expression after transfection in CV-1 green monkey kidney cells. Analysis of multiple 5′ deletion mutants reveals that a minimum of 85 bases upstream of the major transcriptional initiation site are required for full basal promoter activity. Deleting 11 bases to position -74 causes a 4-fold decrease in promoter activity. Another major decrement in activity is seen when a GC box between -46 and -33 is deleted, consistent with this region being a functionally active Sp1 factor-binding site. Primer extension of a CAT-specific primer hybridized to RNA from cells transfected with a Gi2α promoter-CAT construct confirms approximately the same transcriptional start site as the endogenous Gi2α gene. The 3′ deletion mutants that either approach or delete the transcriptional start site have markedly diminished activity.

UR - http://www.scopus.com/inward/record.url?scp=0025109999&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025109999&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 958

EP - 964

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 7

ER -