Thyroid hormone receptor mutations that interfere with transcriptional activation also interfere with receptor interaction with a nuclear protein.
A 20 amino acid segment within the ligand-binding domain of the rat beta 1-thyroid hormone receptor (T3R; amino acids 286-305) is conserved in all members of the erbA superfamily. Mutations in this region impair T3 induction of reporter gene expression in a transfection system without impairing T3 binding or nuclear localization of the receptors. We postulate that a nuclear protein may need to interact with this domain for full transcriptional activity and provide evidence for the existence of this putative protein. Specifically, a JEG-3 cell protein (denoted TRAP, T3R auxiliary protein) interacts with wild-type T3R-beta 1 to increase binding of the receptor to T3 response elements in vitro. Mutations within amino acids 286-305 impair the ability of TRAP to enhance T3R binding to hormone response elements. Cross-linking studies indicate that TRAP has an apparent mol wt of 63 kDa. Surprisingly, TRAP does not enhance binding of erbA-alpha 2 and v-erbA to DNA, even though these proteins contain highly conserved versions of the 20-amino acid beta 1 T3R sequence under study. TRAP or a similar protein, however, does enhance binding of retinoic acid receptor-beta to a hormone response element. We conclude that the inability of T3Rs with mutations in this domain to fully activate transcription may be due to an inability of these proteins to fully interact with TRAP. We postulate that TRAP or related proteins may be important regulators of ligand-dependent transcriptional activation for other members of the erbA family in addition to the T3R.
O'Donnell
A L
AL
Division of Endocrinology, University of Michigan Medical Center, Ann Arbor 48109-0678.
Rosen
E D
ED
Darling
D S
DS
Koenig
R J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D001482:Base Composition; D001665:Binding Sites; D002467:Cell Nucleus / Q000737:chemistry; D004247:DNA / Q000378:metabolism*; D013006:Growth Hormone / Q000235:genetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D046911:Macromolecular Substances; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D011988:Receptors, Thyroid Hormone; D013006:Growth Hormone; D004247:DNA