Regulation of Cre Recombinase Activity by Mutated Estrogen Receptor Ligand-Binding Domains

Biochem Biophys Res Commun. 1997 Aug 28;237(3):752-7. doi: 10.1006/bbrc.1997.7124.

Abstract

Ligand-dependent chimeric Cre recombinases are powerful tools to induce specific DNA rearrangements in cultured cells and in mice. We report here the construction and characterization of a series of chimeric recombinases, each consisting of Cre fused to a mutated human oestrogen receptor (ER) ligand-binding domain (LBD). Two new ligand-dependent recombinases which contain either the G400V/M543A/L544A or the G400V/L539A/L540A triple mutation of the human ER LBD are efficiently induced by the synthetic ER antagonists 4-hydroxytamoxifen (OHT) and ICI 182,780 (ICI), respectively, but are insensitive to 17 beta-oestradiol (E2). Both chimeric recombinases should be useful for efficient spatio-temporally controlled site-directed somatic mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Enzyme Induction / drug effects
  • Estradiol / analogs & derivatives
  • Estradiol / pharmacology
  • Estrogen Antagonists / pharmacology*
  • Fulvestrant
  • Humans
  • Integrases / biosynthesis
  • Integrases / metabolism*
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Mutagenesis, Site-Directed
  • Oligodeoxyribonucleotides
  • Point Mutation
  • Receptors, Estrogen / biosynthesis
  • Receptors, Estrogen / chemistry
  • Receptors, Estrogen / metabolism*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Tamoxifen / analogs & derivatives
  • Tamoxifen / pharmacology
  • Teratoma
  • Tumor Cells, Cultured
  • Viral Proteins*

Substances

  • Estrogen Antagonists
  • Oligodeoxyribonucleotides
  • Receptors, Estrogen
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Tamoxifen
  • afimoxifene
  • Fulvestrant
  • Estradiol
  • Cre recombinase
  • Integrases