Generation of two distinct functional isoforms of dosage-sensitive sex reversal-adrenal hypoplasia congenita-critical region on the X chromosome gene 1 (DAX-1) by alternative splicing

Mol Endocrinol. 2004 Jun;18(6):1428-37. doi: 10.1210/me.2003-0176. Epub 2004 Mar 25.

Abstract

DAX-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita-critical region on the X chromosome gene 1; NR0B1) is an orphan nuclear receptor that plays an important role in the development and functioning of the adrenal gland and hypothalamic-pituitary gonadal axis. The DAX-1 protein acts as a transcriptional repressor of genes involved in the steroidogenic pathway. We have identified a novel isoform encoded by the known exon 1 of DAX-1 and a previously unrecognized exon (exon 2alpha) that lies within intron 1 of DAX-1. This novel transcript, which we designated DAX-1alpha, is terminated at exon 2alpha; the last 70 amino acids of the C-terminal repressor domain encoded by exon 2 are absent. DAX-1alpha encodes a protein of 401 amino acids; the first 389 amino acids are encoded by exon 1 and the last 12 are encoded by exon 2alpha. Using conventional RT-PCR and real-time RT-PCR analyses, we found that DAX-1alpha is abundantly expressed in the adrenal gland, brain, kidney, ovary, and testis. We also found that DAX-1alpha can bind to steroidogenic factor 1 and to DNA but is unable to repress steroidogenic factor 1-mediated transcriptional activation of the reporter gene and acts as an antagonist of DAX-1 under certain conditions.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alternative Splicing*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • COS Cells
  • Cloning, Molecular
  • DAX-1 Orphan Nuclear Receptor
  • DNA Primers / chemistry
  • DNA, Complementary / metabolism
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism*
  • Dose-Response Relationship, Drug
  • Exons
  • HeLa Cells
  • Homeodomain Proteins
  • Humans
  • Immunoprecipitation
  • Introns
  • Models, Genetic
  • Molecular Sequence Data
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Protein Isoforms
  • Protein Structure, Tertiary
  • RNA, Messenger / metabolism
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Retinoic Acid / chemistry*
  • Receptors, Retinoic Acid / metabolism*
  • Repressor Proteins / chemistry*
  • Repressor Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Steroidogenic Factor 1
  • Steroids / metabolism
  • Tissue Distribution
  • Transcription Factors / metabolism
  • Transcription, Genetic
  • Transfection

Substances

  • DAX-1 Orphan Nuclear Receptor
  • DNA Primers
  • DNA, Complementary
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • NR0B1 protein, human
  • NR5A1 protein, human
  • Protein Isoforms
  • RNA, Messenger
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Retinoic Acid
  • Repressor Proteins
  • Steroidogenic Factor 1
  • Steroids
  • Transcription Factors