Binding of MCF-7 cell mitochondrial proteins and recombinant human estrogen receptors α and β to human mitochondrial DNA estrogen response elements

Jin Q. Chen, Matthewos Eshete, William L. Alworth, James D. Yager

Research output: Contribution to journalArticlepeer-review

Abstract

Our previous studies have shown that 17β estradiol (E2) enhances the transcript levels of mitochondrial DNA (mtDNA)-encoded genes and mitochondrial respiratory chain (MRC) activity via estrogen receptors (ER). Others have reported the presence of putative estrogen responsive elements (ERE) in human mtDNA (mtEREs) and detection of ERs in mitochondria of rat uterine and ovary cells. Recently, we demonstrated the E2-enhanced mitochondrial localization of ERα and ERβ, and E2-induced mtDNA transcript levels in MCF-7 cells. In this study, we applied electrophoresis mobility shift assays (EMSAs) and surface plasmon resonance (SPR) to determine if mitochondrial extracts, recombinant human ERα (rhERα), and rhERβ interact with mtEREs. Using EMSAs, we observed that ER-containing mitochondrial extracts bound to mtEREs and the binding was enhanced by E2, whereas the binding of mitochondrial proteins from ERβ-deficient cells was almost undetectable. Both rhERα and rhERβ bound to the mtEREs and their binding was altered by their respective antibodies. However, the ERα antibodies did not alter the binding of MCF-7 cell mitochondrial extracts to mtEREs whereas the binding MCF-7 and MDA-MB-231 cell mitochondrial extracts to mtEREs was reduced by ERβ antibody. These results suggest that the mtERE-bound mitochondrial protein is ERβ. Using SPR, we observed the binding of both ERs to mtEREs and that the binding was increased by E2. These results indicate that the mitochondrial ERs can interact with mtEREs and suggest that they may be directly involved in E2 induction of mtDNA transcription.

Original languageEnglish (US)
Pages (from-to)358-373
Number of pages16
JournalJournal of cellular biochemistry
Volume93
Issue number2
DOIs
StatePublished - 2004

Keywords

  • 17β-estradiol
  • ERα and ERβ
  • Estrogen carcinogenesis
  • Estrogen response elements
  • Mitochondrial DNA
  • Mitochondrial DNA transcription
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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