Differential binding of thyroid hormone receptors to mouse glandular kallikrein gene promoters: evidence for multiple binding regions in the mGK-6 gene.
We have used a DNA-cellulose competition assay to investigate the binding of thyroid hormone receptors to fragments of the mouse glandular kallikrein genes and the human and rat GH genes. Nuclear extracts from human lymphoblastoid IM-9 cells were incubated with [125I]tri-iodothyronine [( 125I]T3) and DNA-cellulose. The ability of cloned gene fragments to compete for radiolabelled receptors bound to DNA-cellulose was compared with that of DNA from pBR322. As previously observed, a 900 bp fragment from the human GH gene showed preferential binding to the thyroid hormone receptor. High-affinity binding was observed with a synthetic fragment of the rat GH gene encompassing positions -163 to -192 but not with a similar fragment from positions -224 to -192. Preferential binding was also observed with fragments of the mouse glandular kallikrein gene, mGK-6. Binding to the entire gene and fragments containing 2300 and 776 bp of the promoter region was identical. Detectable but reduced binding was seen with a shorter fragment. These results suggest that the T3 receptor binds to multiple sites within the first 776 bp of the mGK-6 gene promoter. Potential thyroid hormone response elements can be identified within this region of the gene. In contrast, the kallikrein gene mGK-3, which shows a different response to thyroid hormone from that of mGK-6, showed no significant binding in the comparable promoter region.
Ewen Downie Metabolic Unit, Alfred Hospital, Melbourne, Victoria, Australia.
J W
JW
Barlow
L E
LE
Raggatt
C C
CC
Drinkwater
I G
IG
Lyons
R I
RI
Richards
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007610:Kallikreins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D011401:Promoter Regions, Genetic; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014284:Triiodothyronine / Q000378:metabolism
D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine; D013006:Growth Hormone; D004247:DNA; D007610:Kallikreins