Analysis of oestrogen receptor dimerisation using chimeric proteins.
Sequences essential for dimerisation have been identified in the hormone binding domain of the mouse oestrogen receptor by insertional and point mutagenesis and sequence comparisons reveal that equivalent residues may be conserved in other members of the nuclear hormone receptor superfamily. To assess functional compatibility of this region between members of the receptor superfamily, peptide sequences corresponding to the equivalent regions of the human androgen receptor and retinoic acid receptor have been substituted for the dimerisation domain of the mouse oestrogen receptor. The resulting chimeric proteins were analysed for high affinity DNA binding using a gel retardation assay and shown to bind with reduced affinity compared to the wild type oestrogen receptor. The reduction in DNA binding observed may result from the intramolecular incompatibility of functional elements within the hormone binding domain of nuclear hormone receptors.
White
R
R
Molecular Endocrinology Department, Imperial Cancer Research Fund, London, England.
Fawell
S E
SE
Parker
M G
MG
D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000378:metabolism; D006728:Hormones / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D014176:Protein Biosynthesis; D012313:RNA / Q000096:biosynthesis; D018075:RNA, Complementary; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D006728:Hormones; D018075:RNA, Complementary; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D012313:RNA; D004247:DNA