Thyroid hormone action in the absence of thyroid hormone receptor DNA-binding in vivo
J. Clin. Invest. Nobuyuki Shibusawa, et al. 112:588 doi:10.1172/JCI18377 [Go to this article.]

Figure 2
(a) The amino acid sequence of the DNA-binding domain of TR-β. The boxed region outlines the DNA recognition α-helix. The shaded circles indicate the P-box amino acids. The exchange of glutamic acid 125 and glycine 126 to glycine and serine in the GS125 mutation is indicated. (b) Liver RNA was amplified by RT-PCR using primers located in sequences corresponding to exons 2 and 4 (A-1, A-2) and exons 3 and 4 (B-1, B-2), which are indicated by arrows. (c) RT-PCR results from liver RNA using the indicate primers. A 445-bp fragment from WT (+/+) and TR-β^GS/GS animals was observed with the A primer set. A short fragment (344 bp) from the A primer set and no band from the B set were obtained from RNA transcripts from TR-β^–/– mice. (d) DNA sequence of the 445-bp RT-PCR fragment. The two mutated amino acids are indicated. The deleted mRNA from TR-β^–/– encoded a putative peptide that terminated after eight bases in exon 4 at a TAG stop codon. This was confirmed by sequencing the 344-bp fragment from TR-β^–/– mice (data not shown). (e) Western blot analysis of liver total cellular protein extracts from WT, TR-β^–/–, and TR-β^GS/GS animals (a C-terminal TR-β monoclonal antibody was used), indicating that the GS125 mutation did not affect expression from the TR-β locus.