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GAL4-VP16 Stimulates Two RNA Polymerase II Promoters Primarily at the Preinitiation Complex Assembly Step

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We previously demonstrated that RNA polymerase II promoters may be limited in strength not only at the step of transcription complex assembly, but also at initiation or promoter clearance. Here we report on experiments designed to test the possibility that steps following transcription complex assembly might be stimulated by transcriptional activators. Using an in vitro system in which we can independently measure the efficiency of assembly, initiation, and promoter clearance, we have investigated the mechanism by which the model activator GAL4-VP16 increases transcription from two promoters: a weak variant of Ad 2 ML with an altered TATA box, which is inefficient in transcription initiation, and the mouse β-globin promoter, which is inefficient in promoter clearance. We found that whereas GAL4-VP16 is effective in stimulating both promoters, this increase resulted only from greater transcription complex assembly; the initiation and clearance steps were not affected. Because recent studies have suggested that the core transcription factors TFIIE and TFIIH might be important in promoter clearance, we also attempted to increase the initiation and clearance efficiencies of the Ad ML-TATA mutant and globin promoters by direct addition of excess TFIIE and TFIIH to partially purified preinitiation complexes assembled at each of these promoters. These factors had no effect on transcription by either of the preinitiation complexes.
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Keywords: Complex assembly; Initiation; Promoter clearance; RNA polymerase II promoters; Transcriptional activators

Document Type: Research Article

Publication date: January 1, 1996

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  • Gene Expression, The Journal of Liver Research will publish articles in all aspects of hepatology. Hepatology, as a research discipline, has seen unprecedented growth especially in the cellular and molecular mechanisms of hepatic health and disease, which continues to have a major impact on understanding liver development, stem cells, carcinogenesis, tissue engineering, injury, repair, regeneration, immunology, metabolism, fibrosis, and transplantation. Continued research and improved understanding in these areas will have a meaningful impact on liver disease prevention, diagnosis, and treatment. The existing journal Gene Expression has expanded its focus to become Gene Expression, The Journal of Liver Research to meet this growing demand. In its revised and expanded scope, the journal will publish high-impact original articles, reviews, short but complete articles, and special articles (editorials, commentaries, opinions) on all aspects of hepatology, making it a unique and invaluable resource for readers interested in this field. The expanded team, led by an Editor-in-Chief who is uniquely qualified and a renowned expert, along with a dynamic and functional editorial board, is determined to make this a premier journal in the field of hepatology.
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