Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor-enriched S-49 mouse lymphoma cells.
A cDNA library from plasma membrane glucocorticoid receptor-enriched (mGR(++)) S- 49 mouse T lymphoma cells was screened with full-length rat intracellular GR (iGR) cDNA, BUGR-2 antibody, and PCR amplimers to portions of the mouse GR cDNA. One or two single-base substitutions resulting in amino acid changes (which do not incapacitate the receptor) were found in all but one clone: Val437 --> Gly (located in the first zinc finger), and Glu546 --> Gly (in the steroid-binding domain). Two previously unidentified exon 1 variants (1D and 1E), and two of three previously reported variants (1A, 1B) were found to be spliced onto the common exon 2. Exon 1D- and 1E-containing transcripts were confirmed by direct sequencing of amplimers from reverse transcriptase-coupled PCR. RNase protection studies revealed that one of these transcripts was expressed in mGR(++) cells only, but not in two mGR-less (mGR(--) S-49, and AtT-20 mouse pituitary) cell lines. These studies suggest that at least four promoters may be responsible for the control of GR (iGR and mGR) types in mouse lymphoma cells.
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
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F
Chen
C S
CS
Watson
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B
Gametchu
D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000032:analysis; D005091:Exons; D015723:Gene Library; D008223:Lymphoma / Q000235:genetics* / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010911:Pituitary Neoplasms / Q000378:metabolism; D017354:Point Mutation; D012313:RNA / Q000032:analysis; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D008565:Membrane Proteins; D011965:Receptors, Glucocorticoid; D012313:RNA