In vivo expression of rat liver c-erbA beta thyroid hormone receptor in yeast (Saccharomyces cerevisiae).
To study thyroid hormone receptor (TR), we developed an in vivo expression system in yeast by using a copper-responsive yeast metallothionein promoter and ubiquitin-fusion protein technology. The cDNA encoding full-length rat liver TR beta was expressed under the control of copper. The [125I]T3 binding activities to yeast extracts were significantly correlated with the added copper sulfate into the medium. Partially purified TR from the transformed yeast had a high hormone binding affinity (Kd = 0.34) for T3 and could bind thyroid hormone response element in gel retardation analysis.
Thyroid Research Laboratory, Samuel Lunenfeld Research Institute of Mt. Sinai Hospital, University of Toronto, Ontario, Canada.
C
C
Lu
Y F
YF
Yang
H
H
Ohashi
P G
PG
Walfish
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008099:Liver / Q000502:physiology; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D012045:Regulatory Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014284:Triiodothyronine / Q000378:metabolism
D009841:Oligonucleotides; D011988:Receptors, Thyroid Hormone; D011994:Recombinant Proteins; D014284:Triiodothyronine; D004247:DNA