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J Biol Chem, Vol. 274, Issue 20, 13768-13776, May 14, 1999
From the The type 2 iodothyronine deiodinase (D2) is
critical for the intracellular production of 3,5,3'-triiodothyronine
from thyroxine. The D2 mRNA of higher vertebrates is over 6 kilobases (kb), and no complete cDNA clones have been reported.
Using 5'- and 3'-rapid amplification of cDNA ends and two cDNA
libraries, we have cloned the 6094-base pair full-length chicken D2
cDNA. The deduced protein is ~31 kDa and contains two in-frame
UGA codons presumably encoding selenocysteine. One of these is in the
highly conserved active catalytic center; the other is near the
carboxyl terminus. Unusual features of the cDNA include a
selenocysteine insertion sequence element ~4.8 kb 3' to the UGA codon
in the active center and three short open reading frames in the
5'-untranslated region. The Km of D2 is ~1.0
nM for thyroxine, and the reaction is insensitive to
inhibition by 6-n-propylthiouracil. Chicken D2 is expressed as a single transcript of ~6 kb in different brain regions and in the
thyroid and lung. Hypothyroidism increases D2 mRNA in the telencephalon. Unlike in mammals, D2 mRNA and activity are
expressed in the liver of the chicken, suggesting a role for D2 in the
generation of plasma 3,5,3'-triiodothyronine in this species.
Cloning and Expression of the Chicken Type 2 Iodothyronine
5'-Deiodinase
§,,,
Thyroid Division, Department of Medicine,
Brigham and Women's Hospital and Harvard Medical School, Boston,
Massachusetts 02115 and the § Department of Physiology and
Biochemistry, University of Veterinary Science, P.O. Box 2, Budapest H-1400, Hungary
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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