Activation of cAMP-dependent protein kinase increases the protein level of steroidogenic factor-1

Reidun ÆSøY, Gunnar Mellgren, Ken-Ichirou Morohashi, Johan Lund

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

The orphan nuclear receptor steroidogenic factor 1 (SF-1) is an essential regulator of endocrine organogenesis, sexual differentiation, and steroidogenisis. SF-1 is a transcriptional regulator of cAMP responsive genes, but the exact mechanisms by which cAMP-dependent PKA modulates SF-1 dependent transcription leading to increased steroidogenic output have not been determined. In this report the effects of PKA activation on SF-1 in living cells have been examined by the use of full-length SF-1 cDNA fused to the cDNA encoding green fluorescent protein (GFP). The GFP-SF-1 fusion protein localized to the nucleus of both steroidogenic Y1 cells and non-steroidogenic COS-1 cells, and the functional properties of wild-type SF-1 were conserved. When the catalytic subunit of PKA was coexpressed with GFP-SF-1, we observed that the fluorescence emission was markedly elevated. These findings were confirmed by Western blot analysis, showing that stimulation of PKA increased SF-1 protein levels. The PKA-induced expression of SF-1 protein was not accompanied by an increase in SF-1 mRNA levels. However, pulse-chase studies showed a decrease in SF-1 degradation rate in response to activation of PKA, indicating that PKA elevates the level of SF-1 by increasing the stability of SF-1 protein.

Original languageEnglish
Pages (from-to)295-303
Number of pages9
JournalEndocrinology
Volume143
Issue number1
DOIs
Publication statusPublished - Jan 22 2002

Fingerprint

Steroidogenic Factor 1
Cyclic AMP-Dependent Protein Kinases
Proteins
Green Fluorescent Proteins
Complementary DNA
Orphan Nuclear Receptors
Sex Differentiation
Organogenesis
COS Cells

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Activation of cAMP-dependent protein kinase increases the protein level of steroidogenic factor-1. / ÆSøY, Reidun; Mellgren, Gunnar; Morohashi, Ken-Ichirou; Lund, Johan.

In: Endocrinology, Vol. 143, No. 1, 22.01.2002, p. 295-303.

Research output: Contribution to journalArticle

ÆSøY, Reidun ; Mellgren, Gunnar ; Morohashi, Ken-Ichirou ; Lund, Johan. / Activation of cAMP-dependent protein kinase increases the protein level of steroidogenic factor-1. In: Endocrinology. 2002 ; Vol. 143, No. 1. pp. 295-303.
@article{52db3bbb04f6497ebf057d25a8111b59,
title = "Activation of cAMP-dependent protein kinase increases the protein level of steroidogenic factor-1",
abstract = "The orphan nuclear receptor steroidogenic factor 1 (SF-1) is an essential regulator of endocrine organogenesis, sexual differentiation, and steroidogenisis. SF-1 is a transcriptional regulator of cAMP responsive genes, but the exact mechanisms by which cAMP-dependent PKA modulates SF-1 dependent transcription leading to increased steroidogenic output have not been determined. In this report the effects of PKA activation on SF-1 in living cells have been examined by the use of full-length SF-1 cDNA fused to the cDNA encoding green fluorescent protein (GFP). The GFP-SF-1 fusion protein localized to the nucleus of both steroidogenic Y1 cells and non-steroidogenic COS-1 cells, and the functional properties of wild-type SF-1 were conserved. When the catalytic subunit of PKA was coexpressed with GFP-SF-1, we observed that the fluorescence emission was markedly elevated. These findings were confirmed by Western blot analysis, showing that stimulation of PKA increased SF-1 protein levels. The PKA-induced expression of SF-1 protein was not accompanied by an increase in SF-1 mRNA levels. However, pulse-chase studies showed a decrease in SF-1 degradation rate in response to activation of PKA, indicating that PKA elevates the level of SF-1 by increasing the stability of SF-1 protein.",
author = "Reidun {\AE}S{\o}Y and Gunnar Mellgren and Ken-Ichirou Morohashi and Johan Lund",
year = "2002",
month = "1",
day = "22",
doi = "10.1210/en.143.1.295",
language = "English",
volume = "143",
pages = "295--303",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Activation of cAMP-dependent protein kinase increases the protein level of steroidogenic factor-1

AU - ÆSøY, Reidun

AU - Mellgren, Gunnar

AU - Morohashi, Ken-Ichirou

AU - Lund, Johan

PY - 2002/1/22

Y1 - 2002/1/22

N2 - The orphan nuclear receptor steroidogenic factor 1 (SF-1) is an essential regulator of endocrine organogenesis, sexual differentiation, and steroidogenisis. SF-1 is a transcriptional regulator of cAMP responsive genes, but the exact mechanisms by which cAMP-dependent PKA modulates SF-1 dependent transcription leading to increased steroidogenic output have not been determined. In this report the effects of PKA activation on SF-1 in living cells have been examined by the use of full-length SF-1 cDNA fused to the cDNA encoding green fluorescent protein (GFP). The GFP-SF-1 fusion protein localized to the nucleus of both steroidogenic Y1 cells and non-steroidogenic COS-1 cells, and the functional properties of wild-type SF-1 were conserved. When the catalytic subunit of PKA was coexpressed with GFP-SF-1, we observed that the fluorescence emission was markedly elevated. These findings were confirmed by Western blot analysis, showing that stimulation of PKA increased SF-1 protein levels. The PKA-induced expression of SF-1 protein was not accompanied by an increase in SF-1 mRNA levels. However, pulse-chase studies showed a decrease in SF-1 degradation rate in response to activation of PKA, indicating that PKA elevates the level of SF-1 by increasing the stability of SF-1 protein.

AB - The orphan nuclear receptor steroidogenic factor 1 (SF-1) is an essential regulator of endocrine organogenesis, sexual differentiation, and steroidogenisis. SF-1 is a transcriptional regulator of cAMP responsive genes, but the exact mechanisms by which cAMP-dependent PKA modulates SF-1 dependent transcription leading to increased steroidogenic output have not been determined. In this report the effects of PKA activation on SF-1 in living cells have been examined by the use of full-length SF-1 cDNA fused to the cDNA encoding green fluorescent protein (GFP). The GFP-SF-1 fusion protein localized to the nucleus of both steroidogenic Y1 cells and non-steroidogenic COS-1 cells, and the functional properties of wild-type SF-1 were conserved. When the catalytic subunit of PKA was coexpressed with GFP-SF-1, we observed that the fluorescence emission was markedly elevated. These findings were confirmed by Western blot analysis, showing that stimulation of PKA increased SF-1 protein levels. The PKA-induced expression of SF-1 protein was not accompanied by an increase in SF-1 mRNA levels. However, pulse-chase studies showed a decrease in SF-1 degradation rate in response to activation of PKA, indicating that PKA elevates the level of SF-1 by increasing the stability of SF-1 protein.

UR - http://www.scopus.com/inward/record.url?scp=0000638563&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000638563&partnerID=8YFLogxK

U2 - 10.1210/en.143.1.295

DO - 10.1210/en.143.1.295

M3 - Article

C2 - 11751621

AN - SCOPUS:0000638563

VL - 143

SP - 295

EP - 303

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 1

ER -