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We have investigated the regulation of glutamine synthetase (GS) mRNA synthesis in Chinese hamster ovary cell mutants which overproduce GS and contain an amplified GS gene. Specific mRNA synthesis was analyzed by measuring elongation of transcripts in isolated nuclei. Transcription was assayed by hybridization of newly synthesized [32P]RNA to a genomic GS clone. Nuclear transcript elongation was inhibited more than 90% by alpha-amanitin. The relative rates of GS mRNA synthesis in nuclei from cells incubated for 2 days with no additions, insulin, dexamethasone, or (Bu)2cAMP are 186, 419, 375, and 227 ppm, respectively. The insulin- and dexamethasone-mediated increases in GS transcription rate (2-fold) were associated with 3.7- and 5.8-fold increases in GS mRNA abundance. By contrast, alpha-tubulin gene transcription was not altered by insulin or dexamethasone; however, it was decreased by (Bu)2cAMP.

Insulin and Dexamethasone Stimulate Transcription of an Amplified Glutamine Synthetase Gene in Chinese Hamster Ovary Cells