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Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor‐enriched S‐49 mouse lymphoma cells
Author(s) -
Chen Fanghong,
Watson Cheryl S.,
Gametchu Bahiru
Publication year - 1999
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19990901)74:3<418::aid-jcb10>3.0.co;2-6
Subject(s) - glucocorticoid receptor , microbiology and biotechnology , complementary dna , biology , exon , cdna library , receptor , clone (java method) , gene , biochemistry
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. J. Cell. Biochem. 74:418–429, 1999. © 1999 Wiley‐Liss, Inc.