The DNA binding domain of retinoic acid receptor beta is required for ligand-dependent suppression of proliferation. Application of general purpose mammalian coexpression vectors

J Cell Sci. 1994 Apr;107 ( Pt 4):827-38.

Abstract

We have developed a family of mammalian coexpression vectors that permit identification of living or fixed cells overexpressing a gene of interest by surrogate detection of a coexpressed marker protein. Using these 'pMARK' vectors, a fluorescence-based, single cell proliferation assay was developed and used to study the effect of retinoic acid receptor beta (RAR-beta) on cell cycling. We demonstrate that transient overexpression of RAR-beta in the presence, but not absence, of all-trans retinoic acid results in a dramatic suppression of cell proliferation. We further show that this effect requires the DNA binding (C) domain of RAR-beta. It has been previously shown that RAR-beta expression is markedly altered in a variety of neoplasms and cell lines. Our data support the hypothesis that loss of RAR-beta may contribute to tumor progression by removing normal restraints on proliferation. The pMARK vectors should be useful for studying other genes that putatively suppress or enhance proliferation.

Publication types

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

MeSH terms

  • Base Sequence
  • Binding Sites
  • Cell Division / drug effects
  • Cell Division / physiology*
  • DNA / metabolism*
  • Genetic Vectors*
  • HeLa Cells
  • Humans
  • Ligands
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Secondary*
  • Receptors, Retinoic Acid / chemistry
  • Receptors, Retinoic Acid / genetics
  • Receptors, Retinoic Acid / physiology*
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / biosynthesis
  • Sensitivity and Specificity
  • Transfection
  • Tretinoin / pharmacology

Substances

  • Ligands
  • Receptors, Retinoic Acid
  • Recombinant Fusion Proteins
  • retinoic acid receptor beta
  • Tretinoin
  • DNA