Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor-enriched S-49 mouse lymphoma cells

✍ Scribed by Fanghong Chen; Cheryl S. Watson; Bahiru Gametchu

• Book ID: 101260540
• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 232 KB
• Volume: 74
• Category: Article
• ISSN: 0730-2312
• DOI: 10.1002/(sici)1097-4644(19990901)74:3<418::aid-jcb10>3.0.co;2-6

✦ Synopsis

A cDNA library from plasma membrane glucocorticoid receptor-enriched (mGR ϩϩ ) 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 ϩϩ cells only, but not in two mGR-less (mGR ϪϪ 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.