Analysis of DNA sequences required for pituitary-specific expression of the glycoprotein hormone α-subunit gene

William E. Schoderbek, Kyoon E. Kim, E. Chester Ridgway, Pamela L. Mellon, Richard Maurer

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Transient transfection studies have been used to determine the DNA sequences of the glycoprotein hormone α-subunit gene that are required for tissue-specific expression. In the initial phase of these studies, a variant mouse α gene was identified which contains a fully palindromic cAMP response element (CRE). The corresponding region of a previously cloned and sequenced mouse α gene contains a single point mutation that disrupts the symmetrical nature of this element. DNase footprint studies demonstrate that the fully palindromic CRE binds the CRE-binding protein with much higher affinity than the imperfect palindrome. Transfection experiments using both mouse α gene variants demonstrate differences in basal and cAMP-induced expression. Studies of the cAMP response of the human α gene indicated that this gene contains sequences other than the known CRE that are sufficient to permit a transcriptional response to cAMP in both placental and pituitary cells. Expression of human and mouse α-subunit genes has been examined in cells of the gonadotrope, thyrotrope, and trophoblast lineages to identify DNA sequences that mediate selective transcription of the α gene in these cells. The results demonstrate that sequences between about -500 and -200 are important for expression in the pituitary, but not the placenta. Clustered point mutations were used to further characterize sequences required for expression in the pituitary. Two regions, one at positions -445 to -438 and one at positions -337 to -330, were required for expression in cells of the gonadotrope lineage. One of these regions, at -337 to -330, is also important for expression in thyrotropes. When linked to a minimal promoter, multiple copies of the -344 to -300 region had transcriptional enhancer activity in gonadotropes and thyrotropes, but not in several other cell types. These results are consistent with a model involving different combinations of regulatory elements that determine cell-specific α expression in gonadotropes and thyrotropes.

Original languageEnglish (US)
Pages (from-to)893-903
Number of pages11
JournalMolecular Endocrinology
Volume6
Issue number6
StatePublished - Jun 1992
Externally publishedYes

Fingerprint

DNA Sequence Analysis
Glycoproteins
Hormones
Genes
Response Elements
Point Mutation
Transfection
Cyclic AMP Response Element-Binding Protein
Deoxyribonucleases
Trophoblasts
Cell Lineage
Placenta

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Analysis of DNA sequences required for pituitary-specific expression of the glycoprotein hormone α-subunit gene. / Schoderbek, William E.; Kim, Kyoon E.; Ridgway, E. Chester; Mellon, Pamela L.; Maurer, Richard.

In: Molecular Endocrinology, Vol. 6, No. 6, 06.1992, p. 893-903.

Research output: Contribution to journalArticle

Schoderbek, William E. ; Kim, Kyoon E. ; Ridgway, E. Chester ; Mellon, Pamela L. ; Maurer, Richard. / Analysis of DNA sequences required for pituitary-specific expression of the glycoprotein hormone α-subunit gene. In: Molecular Endocrinology. 1992 ; Vol. 6, No. 6. pp. 893-903.
@article{c3d7d9257b04462d8eb563104384eae3,
title = "Analysis of DNA sequences required for pituitary-specific expression of the glycoprotein hormone α-subunit gene",
abstract = "Transient transfection studies have been used to determine the DNA sequences of the glycoprotein hormone α-subunit gene that are required for tissue-specific expression. In the initial phase of these studies, a variant mouse α gene was identified which contains a fully palindromic cAMP response element (CRE). The corresponding region of a previously cloned and sequenced mouse α gene contains a single point mutation that disrupts the symmetrical nature of this element. DNase footprint studies demonstrate that the fully palindromic CRE binds the CRE-binding protein with much higher affinity than the imperfect palindrome. Transfection experiments using both mouse α gene variants demonstrate differences in basal and cAMP-induced expression. Studies of the cAMP response of the human α gene indicated that this gene contains sequences other than the known CRE that are sufficient to permit a transcriptional response to cAMP in both placental and pituitary cells. Expression of human and mouse α-subunit genes has been examined in cells of the gonadotrope, thyrotrope, and trophoblast lineages to identify DNA sequences that mediate selective transcription of the α gene in these cells. The results demonstrate that sequences between about -500 and -200 are important for expression in the pituitary, but not the placenta. Clustered point mutations were used to further characterize sequences required for expression in the pituitary. Two regions, one at positions -445 to -438 and one at positions -337 to -330, were required for expression in cells of the gonadotrope lineage. One of these regions, at -337 to -330, is also important for expression in thyrotropes. When linked to a minimal promoter, multiple copies of the -344 to -300 region had transcriptional enhancer activity in gonadotropes and thyrotropes, but not in several other cell types. These results are consistent with a model involving different combinations of regulatory elements that determine cell-specific α expression in gonadotropes and thyrotropes.",
author = "Schoderbek, {William E.} and Kim, {Kyoon E.} and Ridgway, {E. Chester} and Mellon, {Pamela L.} and Richard Maurer",
year = "1992",
month = "6",
language = "English (US)",
volume = "6",
pages = "893--903",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "6",

}

TY - JOUR

T1 - Analysis of DNA sequences required for pituitary-specific expression of the glycoprotein hormone α-subunit gene

AU - Schoderbek, William E.

AU - Kim, Kyoon E.

AU - Ridgway, E. Chester

AU - Mellon, Pamela L.

AU - Maurer, Richard

PY - 1992/6

Y1 - 1992/6

N2 - Transient transfection studies have been used to determine the DNA sequences of the glycoprotein hormone α-subunit gene that are required for tissue-specific expression. In the initial phase of these studies, a variant mouse α gene was identified which contains a fully palindromic cAMP response element (CRE). The corresponding region of a previously cloned and sequenced mouse α gene contains a single point mutation that disrupts the symmetrical nature of this element. DNase footprint studies demonstrate that the fully palindromic CRE binds the CRE-binding protein with much higher affinity than the imperfect palindrome. Transfection experiments using both mouse α gene variants demonstrate differences in basal and cAMP-induced expression. Studies of the cAMP response of the human α gene indicated that this gene contains sequences other than the known CRE that are sufficient to permit a transcriptional response to cAMP in both placental and pituitary cells. Expression of human and mouse α-subunit genes has been examined in cells of the gonadotrope, thyrotrope, and trophoblast lineages to identify DNA sequences that mediate selective transcription of the α gene in these cells. The results demonstrate that sequences between about -500 and -200 are important for expression in the pituitary, but not the placenta. Clustered point mutations were used to further characterize sequences required for expression in the pituitary. Two regions, one at positions -445 to -438 and one at positions -337 to -330, were required for expression in cells of the gonadotrope lineage. One of these regions, at -337 to -330, is also important for expression in thyrotropes. When linked to a minimal promoter, multiple copies of the -344 to -300 region had transcriptional enhancer activity in gonadotropes and thyrotropes, but not in several other cell types. These results are consistent with a model involving different combinations of regulatory elements that determine cell-specific α expression in gonadotropes and thyrotropes.

AB - Transient transfection studies have been used to determine the DNA sequences of the glycoprotein hormone α-subunit gene that are required for tissue-specific expression. In the initial phase of these studies, a variant mouse α gene was identified which contains a fully palindromic cAMP response element (CRE). The corresponding region of a previously cloned and sequenced mouse α gene contains a single point mutation that disrupts the symmetrical nature of this element. DNase footprint studies demonstrate that the fully palindromic CRE binds the CRE-binding protein with much higher affinity than the imperfect palindrome. Transfection experiments using both mouse α gene variants demonstrate differences in basal and cAMP-induced expression. Studies of the cAMP response of the human α gene indicated that this gene contains sequences other than the known CRE that are sufficient to permit a transcriptional response to cAMP in both placental and pituitary cells. Expression of human and mouse α-subunit genes has been examined in cells of the gonadotrope, thyrotrope, and trophoblast lineages to identify DNA sequences that mediate selective transcription of the α gene in these cells. The results demonstrate that sequences between about -500 and -200 are important for expression in the pituitary, but not the placenta. Clustered point mutations were used to further characterize sequences required for expression in the pituitary. Two regions, one at positions -445 to -438 and one at positions -337 to -330, were required for expression in cells of the gonadotrope lineage. One of these regions, at -337 to -330, is also important for expression in thyrotropes. When linked to a minimal promoter, multiple copies of the -344 to -300 region had transcriptional enhancer activity in gonadotropes and thyrotropes, but not in several other cell types. These results are consistent with a model involving different combinations of regulatory elements that determine cell-specific α expression in gonadotropes and thyrotropes.

UR - http://www.scopus.com/inward/record.url?scp=0026752378&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026752378&partnerID=8YFLogxK

M3 - Article

VL - 6

SP - 893

EP - 903

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 6

ER -