The in vitro transcription of the 7SK RNA gene by RNA polymerase III is dependent only on the presence of an upstream promoter

• Published in: Cell Vol. 51; no. 1; p. 81
• Main Authors: Murphy, S, Di Liegro, C, Melli, M
• Format: Journal Article
• Language: English
• Published: United States 09-10-1987
• Subjects: Amanitins - pharmacology; Base Sequence; DNA, Recombinant; DNA-Directed RNA Polymerases - metabolism; Electrophoresis, Polyacrylamide Gel; Humans; Molecular Sequence Data; Mutation; Plasmids; Promoter Regions, Genetic; Regulatory Sequences, Nucleic Acid; RNA Polymerase III - metabolism; RNA, Small Nuclear - genetics; Transcription, Genetic - drug effects
• ISSN: 0092-8674
• DOI: 10.1016/0092-8674(87)90012-2

Deletion analysis was carried out on the human 7SK RNA gene to map regions essential for in vitro transcription by RNA polymerase III. The sequence promoting transcription is located between 37 and 3 bp upstream of the 7SK RNA coding region. RNA polymerase III transcription of adjacent plasmid sequences can be directed by this promoter in the complete absence of the 7SK RNA coding region, indicating that no internal promoter sequences are required. Transcription is terminated by a stretch of T residues, typical of RNA polymerase III transcription. The promoter contains a TATA box at position -25, mutations within which dramatically reduce the efficiency of transcription. Upstream sequences from position -37 to -243 increase the promoter's efficiency. The promoter recognized by RNA polymerase III is structurally and functionally similar to the promoter of genes transcribed by RNA polymerase II.