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FOXL2 interacts with steroidogenic factor-1 (SF-1) and represses SF-1-induced CYP17 transcription in granulosa cells

Mira Park1, Eunkyoung Shin, Miae Won

  • 1Department of Biomedical Science, College of Life Science, CHA University, Seongnam 463-836, Korea.

Molecular Endocrinology (Baltimore, Md.)
|March 9, 2010
PubMed

Insights

Mutations in FOXL2 cause blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) and ovarian failure. This study reveals FOXL2 normally inhibits SF-1

Area of Science:

  • Reproductive Biology
  • Endocrinology
  • Genetics

Background:

  • Mutations in the FOXL2 gene are linked to blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) type I, characterized by premature ovarian failure in affected women.
  • FOXL2 plays a crucial role in ovarian granulosa cell development and function during folliculogenesis, as evidenced by defects in FOXL2-null mice.

Purpose of the Study:

  • To identify proteins interacting with FOXL2 in ovarian cells.
  • To elucidate the regulatory mechanism of FOXL2 on steroidogenesis, particularly concerning the CYP17 gene.
  • To investigate how FOXL2 mutations associated with BPES type I affect its interaction with SF-1 and regulation of CYP17.

Main Methods:

  • Screening of a rat ovarian yeast two-hybrid cDNA library to identify FOXL2-interacting proteins.
  • Co-immunoprecipitation assays to confirm the interaction between human FOXL2 and SF-1 in human granulosa cells.
  • Reporter gene assays to assess the regulation of CYP17 transcriptional activation by FOXL2 and SF-1.
  • Chromatin immunoprecipitation (ChIP) and Electrophoretic Mobility Shift Assay (EMSA) to analyze protein-DNA interactions at the CYP17 promoter.

Main Results:

  • Steroidogenic factor-1 (SF-1) was identified as a FOXL2-interacting protein.
  • FOXL2 negatively regulates SF-1-mediated transcriptional activation of the CYP17 gene in human granulosa cells.
  • FOXL2 mutants associated with BPES type I lose the ability to repress SF-1-induced CYP17 expression.
  • FOXL2 directly inhibits the binding of SF-1 to the CYP17 promoter, a function impaired in BPES-associated mutants.

Conclusions:

  • This study identifies a novel role for FOXL2 in negatively regulating the expression of the key steroidogenic enzyme CYP17 via interaction with SF-1.
  • Impaired inhibition of SF-1 binding to the CYP17 promoter by FOXL2 mutants provides a molecular mechanism for ovarian dysfunction in BPES type I.
  • FOXL2's regulation of CYP17 is critical for normal ovarian follicle development and function.

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