Identification of C. elegans DAF-12-binding sites, response elements, and target genes

Genes Dev. 2004 Oct 15;18(20):2529-44. doi: 10.1101/gad.1218504.

Abstract

Intracellular receptor DAF-12 regulates dauer formation and developmental age and affects Caenorhabditis elegans lifespan. Genetic analyses place DAF-12 at the convergence of several signal transduction pathways; however, the downstream effectors and the molecular basis for the receptor's multiple physiological outputs are unknown. Beginning with C. elegans genomic DNA, we devised a procedure for multiple rounds of selection and amplification that yielded fragments bearing DAF-12-binding sites. These genomic fragments mediated DAF-12-dependent transcriptional regulation both in Saccharomyces cerevisiae and in C. elegans; that is, they served as functional DAF-12 response elements. We determined that most of the genomic fragments that displayed DAF-12 response element activity in yeast were linked to genes that were regulated by DAF-12 in C. elegans; indeed, the response element-containing fragments typically resided within clusters of DAF-12-regulated genes. DAF-12 target gene regulation was developmental program and stage specific, potentially predicting a fit of these targets into regulatory networks governing aspects of C. elegans reproductive development and dauer formation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites / physiology
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism*
  • Caenorhabditis elegans Proteins / physiology
  • Fluorescence Polarization
  • Gene Expression Regulation*
  • Genes, Regulator / genetics
  • Genes, Regulator / physiology*
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins
  • Molecular Sequence Data
  • Oligonucleotides
  • Receptors, Cytoplasmic and Nuclear / genetics*
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Receptors, Cytoplasmic and Nuclear / physiology
  • Response Elements / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Saccharomyces cerevisiae
  • Sequence Analysis, DNA
  • Signal Transduction / physiology

Substances

  • Caenorhabditis elegans Proteins
  • DAF-12 protein, C elegans
  • Oligonucleotides
  • Receptors, Cytoplasmic and Nuclear
  • Green Fluorescent Proteins