Avian and mammalian receptors for 1,25-dihydroxyvitamin D/sub 3/: in vitro translation to characterize size and hormone-dependent regulation

Journal Article · Thu Jan 01 04:00:00 EST 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.2.354· OSTI ID:6125613

Mangelsdorf, D J; Pike, J W; Haussler, M R

In vitro translation of cellular poly(A)/sup +/ RNA coupled with immunoprecipitation was developed as a technique for characterizing 1,25-dihydroxyvitamin D/sub 3/ (1,25-(OH)/sub 2/D/sub 3/) receptors and assessing receptor mRNA activity. Cell-free translation of poly(A)/sup +/ RNA isolated from chicken intestine revealed two immunoprecipitable forms of avian receptor at 60 kDa and 58 kDa. 1,25-(OH)/sub 2/D/sub 3/ receptors were also synthesized in vitro employing poly(A)/sup +/ RNA obtained from several cultured mammalian cell lines. Selective immunoprecipitation revealed a single form of receptor at 54 kDa in mouse fibroblasts (3T6) and pig kidney cells (LLC-PK/sub 1/) and a 52-kDa species in human breast carcinoma (T47D). Each of these in vitro translated mammalian 1,25-(OH)/sub 2/D/sub 3/ receptors migrated identically with its cellular counterpart that was synthesized in vivo employing metabolic labeling of cell protein with (/sup 35/S)methionine. These results are consistent with the conclusions that 1,25-(OH)/sub 2/D/sub 3/ receptors are protein species ranging from 52 to 60 kDa and that, though their functional and immunological domains have been evolutionarily conserved, an inverse relationship apparently exists between phylogenetic status and receptor mass. The data also support the hypothesis that the presence of 1,25-(OH)/sub 2/D/sub 3/ leads to a significant increase in receptor mRNA activity in 3T6 cells, indicative of receptor autoregulation.

• Research Organization: Univ. of Arizona, Tucson
• OSTI ID: 6125613
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:2; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIRDS
BODY
CARBOXYLIC ACIDS
CHICKENS
CHOLECALCIFEROL
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DOMESTIC ANIMALS
DRUGS
ELECTROPHORESIS
EVEN-ODD NUCLEI
FIBROBLASTS
FOWL
GASTROINTESTINAL TRACT
GENE REGULATION
GLANDS
IN VITRO
INTESTINES
ISOTOPES
KIDNEYS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MAMMARY GLANDS
MAN
MEMBRANE PROTEINS
MESSENGER-RNA
METHIONINE
MICE
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PRIMATES
PROTEINS
RADIOISOTOPES
RECEPTORS
RNA
RODENTS
SOMATIC CELLS
SULFUR 35
SULFUR ISOTOPES
SWINE
TUMOR CELLS
VERTEBRATES
VITAMIN D
VITAMINS