Differential regulation of baboon SP-A1 and SP-A2 genes

Structural and functional analysis of 5′-flanking DNA

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Surfactant protein (SP) A gene transcription is developmentally regulated and stimulated by hormones and factors that increase intracellular cAMP. The baboon (b) genome contains two highly similar SP-A genes, bSP-Al and bSP-A2. With the use of a ribonuclease protection assay with gene-specific probes, the two bSP-A genes were found to be differentially regulated during baboon fetal lung development in that expression of the ÖSP-A2 gene appeared to be induced to a high level at a later time in gestation than that of the bSP-Al gene. Both the bSP-Al and bSP-A2 genes were found to be highly responsive to the inductive effects of cAMP in baboon fetal lung expiants in culture. By DNase I footprinting and electrophoretic mobility shift assays with bacterially expressed thyroid transcription factor-1 (TTF-1) and type II cell nuclear extracts, three TTF-1 binding elements were identified within the 255-bp region flanking the 5'-end of each bSP-A gene; however, these differed in position and spacing for the two bSP-A genes. To functionally define the genomic regions that are required for cAMP regulation of bSP-A gene expression in type II cells, fusion genes composed of various amounts of 5′-flanking DNA from the bSP-Al and bSP-A2 genes linked to the human growth hormone structural gene as a reporter were transfected into type II cells in primary culture. We found that 255 bp of 5′-flanking DNA, which contain three TTF-1 binding elements, from bSP-Al and bSP-A2 genes were sufficient to mediate high basal and cAMP-inducible expression in type II cells. We also observed that there were no obvious differences in the magnitude of the responses of these fusion genes to cAMP treatment. surfactant protein A; deoxyribonucleic acid; fetal lung; thyroid transcription factor-1; ribonuclease protection assay

Original languageEnglish (US)
JournalAmerican Journal of Physiology
Volume275
Issue number6 PART 1
StatePublished - 1998

Fingerprint

Papio
varespladib methyl
Surface-Active Agents
DNA
vif Genes
Genes
Proteins
Pulmonary Surfactant-Associated Protein A
Ribonucleases
Lung
Primary Cell Culture
Human Growth Hormone
Cell Fusion
5' Flanking Region
Gene Fusion
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Fetal Development
Cell Extracts
Genome

ASJC Scopus subject areas

  • Physiology (medical)

Cite this

@article{d4148e68156f4a4daab028b6756d4ded,
title = "Differential regulation of baboon SP-A1 and SP-A2 genes: Structural and functional analysis of 5′-flanking DNA",
abstract = "Surfactant protein (SP) A gene transcription is developmentally regulated and stimulated by hormones and factors that increase intracellular cAMP. The baboon (b) genome contains two highly similar SP-A genes, bSP-Al and bSP-A2. With the use of a ribonuclease protection assay with gene-specific probes, the two bSP-A genes were found to be differentially regulated during baboon fetal lung development in that expression of the {\"O}SP-A2 gene appeared to be induced to a high level at a later time in gestation than that of the bSP-Al gene. Both the bSP-Al and bSP-A2 genes were found to be highly responsive to the inductive effects of cAMP in baboon fetal lung expiants in culture. By DNase I footprinting and electrophoretic mobility shift assays with bacterially expressed thyroid transcription factor-1 (TTF-1) and type II cell nuclear extracts, three TTF-1 binding elements were identified within the 255-bp region flanking the 5'-end of each bSP-A gene; however, these differed in position and spacing for the two bSP-A genes. To functionally define the genomic regions that are required for cAMP regulation of bSP-A gene expression in type II cells, fusion genes composed of various amounts of 5′-flanking DNA from the bSP-Al and bSP-A2 genes linked to the human growth hormone structural gene as a reporter were transfected into type II cells in primary culture. We found that 255 bp of 5′-flanking DNA, which contain three TTF-1 binding elements, from bSP-Al and bSP-A2 genes were sufficient to mediate high basal and cAMP-inducible expression in type II cells. We also observed that there were no obvious differences in the magnitude of the responses of these fusion genes to cAMP treatment. surfactant protein A; deoxyribonucleic acid; fetal lung; thyroid transcription factor-1; ribonuclease protection assay",
author = "Mendelson, {C. R.}",
year = "1998",
language = "English (US)",
volume = "275",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "6 PART 1",

}

TY - JOUR

T1 - Differential regulation of baboon SP-A1 and SP-A2 genes

T2 - Structural and functional analysis of 5′-flanking DNA

AU - Mendelson, C. R.

PY - 1998

Y1 - 1998

N2 - Surfactant protein (SP) A gene transcription is developmentally regulated and stimulated by hormones and factors that increase intracellular cAMP. The baboon (b) genome contains two highly similar SP-A genes, bSP-Al and bSP-A2. With the use of a ribonuclease protection assay with gene-specific probes, the two bSP-A genes were found to be differentially regulated during baboon fetal lung development in that expression of the ÖSP-A2 gene appeared to be induced to a high level at a later time in gestation than that of the bSP-Al gene. Both the bSP-Al and bSP-A2 genes were found to be highly responsive to the inductive effects of cAMP in baboon fetal lung expiants in culture. By DNase I footprinting and electrophoretic mobility shift assays with bacterially expressed thyroid transcription factor-1 (TTF-1) and type II cell nuclear extracts, three TTF-1 binding elements were identified within the 255-bp region flanking the 5'-end of each bSP-A gene; however, these differed in position and spacing for the two bSP-A genes. To functionally define the genomic regions that are required for cAMP regulation of bSP-A gene expression in type II cells, fusion genes composed of various amounts of 5′-flanking DNA from the bSP-Al and bSP-A2 genes linked to the human growth hormone structural gene as a reporter were transfected into type II cells in primary culture. We found that 255 bp of 5′-flanking DNA, which contain three TTF-1 binding elements, from bSP-Al and bSP-A2 genes were sufficient to mediate high basal and cAMP-inducible expression in type II cells. We also observed that there were no obvious differences in the magnitude of the responses of these fusion genes to cAMP treatment. surfactant protein A; deoxyribonucleic acid; fetal lung; thyroid transcription factor-1; ribonuclease protection assay

AB - Surfactant protein (SP) A gene transcription is developmentally regulated and stimulated by hormones and factors that increase intracellular cAMP. The baboon (b) genome contains two highly similar SP-A genes, bSP-Al and bSP-A2. With the use of a ribonuclease protection assay with gene-specific probes, the two bSP-A genes were found to be differentially regulated during baboon fetal lung development in that expression of the ÖSP-A2 gene appeared to be induced to a high level at a later time in gestation than that of the bSP-Al gene. Both the bSP-Al and bSP-A2 genes were found to be highly responsive to the inductive effects of cAMP in baboon fetal lung expiants in culture. By DNase I footprinting and electrophoretic mobility shift assays with bacterially expressed thyroid transcription factor-1 (TTF-1) and type II cell nuclear extracts, three TTF-1 binding elements were identified within the 255-bp region flanking the 5'-end of each bSP-A gene; however, these differed in position and spacing for the two bSP-A genes. To functionally define the genomic regions that are required for cAMP regulation of bSP-A gene expression in type II cells, fusion genes composed of various amounts of 5′-flanking DNA from the bSP-Al and bSP-A2 genes linked to the human growth hormone structural gene as a reporter were transfected into type II cells in primary culture. We found that 255 bp of 5′-flanking DNA, which contain three TTF-1 binding elements, from bSP-Al and bSP-A2 genes were sufficient to mediate high basal and cAMP-inducible expression in type II cells. We also observed that there were no obvious differences in the magnitude of the responses of these fusion genes to cAMP treatment. surfactant protein A; deoxyribonucleic acid; fetal lung; thyroid transcription factor-1; ribonuclease protection assay

UR - http://www.scopus.com/inward/record.url?scp=33746722668&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33746722668&partnerID=8YFLogxK

M3 - Article

VL - 275

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 6 PART 1

ER -