A novel A/T-rich element mediates ANF gene expression during cardiac myocyte hypertrophy

Pilar Ruiz-Lozano, Yiu Fai Chen, Paymon Sionit, Yie Teh Yu, Brenda Lilly, Eric N. Olson, Kenneth R. Chien, Adrienne Noel Harris

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

The induction of the atrial natriuretic factor (ANF) gene during α1-adrenergic stimulation of neonatal rat ventricular myocytes has served as a model for gene expression during cardiac muscle cell hypertrophy. This study describes and identifies a single regulatory element that mediates expression of the ANF gene. Deletional mutations were generated in a 639-bp fragment of the ANF promoter that confers α1-adrenergic inducibility to a luciferase reporter gene in transient transfection assays in ventricular myocytes. The results of gel mobility shift and diethylpyrocarbonate (DEPC) interference studies with nuclear cardiac cell extracts identified the nucleotide contract points for a novel A/T-rich element (ANF-AT) at positions -582/-575 that partially mediates α1-adrenergic inducibility. Mutations in the ANF-AT element reduced α1-adrenergic inducibility of an ANF-TK-luciferase fusion gene in cardiac cells by 35%, but had no effect on expression in other muscle and non-muscle cells tested. Gel mobility supershift assays with antibodies directed against the MEF-2 protein, the homeobox protein MHox, or the zinc finger protein HF-1b, document that these factors are not major components of the endogenous ANF-AT binding activity in cardiac muscle cells. The current study provides evidence for a role for a novel A/T-rich element in the regulation of ANF gene expression in cardiac ventricular myocytes.

Original languageEnglish (US)
Pages (from-to)515-525
Number of pages11
JournalJournal of Molecular and Cellular Cardiology
Volume29
Issue number2
DOIs
StatePublished - Feb 1997

    Fingerprint

Keywords

  • A/T-rich binding proteins
  • Atrial natriuretic factor
  • Gene expression
  • Hypertrophy
  • RSRF proteins
  • Transcription
  • α-adrenergic stimulation

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this