The DNA binding domain of retinoic acid receptor beta is required for ligand-dependent suppression of proliferation. Application of general purpose mammalian coexpression vectors.
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.
Frangioni
J V
JV
Molecular Medicine Unit, Beth Israel Hospital, Boston, MA 02215.
Moghal
N
N
Stuart-Tilley
A
A
Neel
B G
BG
Alper
S L
SL
D001483:Base Sequence; D001665:Binding Sites; D002455:Cell Division / Q000187:drug effects / Q000502:physiology*; D004247:DNA / Q000378:metabolism*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary; D018168:Receptors, Retinoic Acid / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D012680:Sensitivity and Specificity; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology
D008024:Ligands; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; C082925:retinoic acid receptor beta; D014212:Tretinoin; D004247:DNA