Isolation and characterization of the 5'-upstream region of the human N- type calcium channel subunit gene

Chromosomal localization and promoter analysis

Dong S. Kim, Hyun Ho Jung, Sun-Hwa Park, Hemin Chin

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

ω-Conotoxin-sensitive N-type Ca2+ channels, unlike dihydropyridine- sensitive L-type channels, are exclusively expressed in nervous tissues. To understand the molecular basis for neuron-specific expression of the N-type channel, we have isolated genomic clones encoding the human α(1B) subunit gene, localized to the long arm of chromosome 9 (9q34) by fluorescence in situ hybridization, and characterized its 5'-upstream region. The proximaI promoter of the α(1B) subunit gene lacks a typical TATA box, is highly GC- rich, and contains several sequences for transcription factor binding. Primer extension experiments revealed the presence of two transcription start sites. In vitro transfection study of the α(1B) subunit-luciferase fusion gene showed that the 4.0-kb 5'-flanking region of the α(1B) gene functions as an efficient promoter in neuronal cells but not in glioma or nonneuronal cells, consistent with the patterns of the endogenous α(1B) gene expression in these cells. Deletion analysis of α(1B) subunit-luciferase fusion gene constructs further revealed the presence of several cis-acting regulatory elements, including a potential repressor located in the distal upstream region (-3992 to -1788) that may be important for the neuron-specific expression of the N-type Ca2+ channel α(1B) subunit gene.

Original languageEnglish
Pages (from-to)5098-5104
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number8
DOIs
Publication statusPublished - 1997 Feb 21

Fingerprint

N-Type Calcium Channels
Genes
Gene Fusion
Luciferases
Conotoxins
Neurons
Nerve Tissue
Chromosomes, Human, Pair 9
TATA Box
5' Flanking Region
Transcription Initiation Site
Fusion reactions
Fluorescence In Situ Hybridization
Glioma
Transfection
Transcription Factors
Clone Cells
Chromosomes
Gene Expression
Gene expression

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Isolation and characterization of the 5'-upstream region of the human N- type calcium channel subunit gene : Chromosomal localization and promoter analysis. / Kim, Dong S.; Jung, Hyun Ho; Park, Sun-Hwa; Chin, Hemin.

In: Journal of Biological Chemistry, Vol. 272, No. 8, 21.02.1997, p. 5098-5104.

Research output: Contribution to journalArticle

@article{a7156db0460d4bf39af6d0feeea228d5,
title = "Isolation and characterization of the 5'-upstream region of the human N- type calcium channel subunit gene: Chromosomal localization and promoter analysis",
abstract = "ω-Conotoxin-sensitive N-type Ca2+ channels, unlike dihydropyridine- sensitive L-type channels, are exclusively expressed in nervous tissues. To understand the molecular basis for neuron-specific expression of the N-type channel, we have isolated genomic clones encoding the human α(1B) subunit gene, localized to the long arm of chromosome 9 (9q34) by fluorescence in situ hybridization, and characterized its 5'-upstream region. The proximaI promoter of the α(1B) subunit gene lacks a typical TATA box, is highly GC- rich, and contains several sequences for transcription factor binding. Primer extension experiments revealed the presence of two transcription start sites. In vitro transfection study of the α(1B) subunit-luciferase fusion gene showed that the 4.0-kb 5'-flanking region of the α(1B) gene functions as an efficient promoter in neuronal cells but not in glioma or nonneuronal cells, consistent with the patterns of the endogenous α(1B) gene expression in these cells. Deletion analysis of α(1B) subunit-luciferase fusion gene constructs further revealed the presence of several cis-acting regulatory elements, including a potential repressor located in the distal upstream region (-3992 to -1788) that may be important for the neuron-specific expression of the N-type Ca2+ channel α(1B) subunit gene.",
author = "Kim, {Dong S.} and Jung, {Hyun Ho} and Sun-Hwa Park and Hemin Chin",
year = "1997",
month = "2",
day = "21",
doi = "10.1074/jbc.272.8.5098",
language = "English",
volume = "272",
pages = "5098--5104",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "8",

}

TY - JOUR

T1 - Isolation and characterization of the 5'-upstream region of the human N- type calcium channel subunit gene

T2 - Chromosomal localization and promoter analysis

AU - Kim, Dong S.

AU - Jung, Hyun Ho

AU - Park, Sun-Hwa

AU - Chin, Hemin

PY - 1997/2/21

Y1 - 1997/2/21

N2 - ω-Conotoxin-sensitive N-type Ca2+ channels, unlike dihydropyridine- sensitive L-type channels, are exclusively expressed in nervous tissues. To understand the molecular basis for neuron-specific expression of the N-type channel, we have isolated genomic clones encoding the human α(1B) subunit gene, localized to the long arm of chromosome 9 (9q34) by fluorescence in situ hybridization, and characterized its 5'-upstream region. The proximaI promoter of the α(1B) subunit gene lacks a typical TATA box, is highly GC- rich, and contains several sequences for transcription factor binding. Primer extension experiments revealed the presence of two transcription start sites. In vitro transfection study of the α(1B) subunit-luciferase fusion gene showed that the 4.0-kb 5'-flanking region of the α(1B) gene functions as an efficient promoter in neuronal cells but not in glioma or nonneuronal cells, consistent with the patterns of the endogenous α(1B) gene expression in these cells. Deletion analysis of α(1B) subunit-luciferase fusion gene constructs further revealed the presence of several cis-acting regulatory elements, including a potential repressor located in the distal upstream region (-3992 to -1788) that may be important for the neuron-specific expression of the N-type Ca2+ channel α(1B) subunit gene.

AB - ω-Conotoxin-sensitive N-type Ca2+ channels, unlike dihydropyridine- sensitive L-type channels, are exclusively expressed in nervous tissues. To understand the molecular basis for neuron-specific expression of the N-type channel, we have isolated genomic clones encoding the human α(1B) subunit gene, localized to the long arm of chromosome 9 (9q34) by fluorescence in situ hybridization, and characterized its 5'-upstream region. The proximaI promoter of the α(1B) subunit gene lacks a typical TATA box, is highly GC- rich, and contains several sequences for transcription factor binding. Primer extension experiments revealed the presence of two transcription start sites. In vitro transfection study of the α(1B) subunit-luciferase fusion gene showed that the 4.0-kb 5'-flanking region of the α(1B) gene functions as an efficient promoter in neuronal cells but not in glioma or nonneuronal cells, consistent with the patterns of the endogenous α(1B) gene expression in these cells. Deletion analysis of α(1B) subunit-luciferase fusion gene constructs further revealed the presence of several cis-acting regulatory elements, including a potential repressor located in the distal upstream region (-3992 to -1788) that may be important for the neuron-specific expression of the N-type Ca2+ channel α(1B) subunit gene.

UR - http://www.scopus.com/inward/record.url?scp=0031053983&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031053983&partnerID=8YFLogxK

U2 - 10.1074/jbc.272.8.5098

DO - 10.1074/jbc.272.8.5098

M3 - Article

VL - 272

SP - 5098

EP - 5104

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 8

ER -