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We have developed a coupled in vitro transcription-polyadenylation system to investigate RNA polymerase II (Pol II) termination, which depends on active polyadenylation of the nascent RNA. Specific G-rich sequences originally identified as binding sites for the transcription factor MAZ both pause Pol II and activate polyadenylation of an upstream poly(A) signal. They do not affect polyadenylation efficiency in an uncoupled cleavage assay. In contrast, pausing of Pol II elongation induced by a high-affinity DNA-binding protein does not activate polyadenylation, indicating that G-rich MAZ sequences have a specific effect on polyadenylation. They also promote intrinsic pausing of purified Pol II, indicating a general role in the modulation of cotranscriptional RNA processing events.

Type

Journal article

Journal

Mol Cell

Publication Date

05/1999

Volume

3

Pages

593 - 600

Keywords

Cell Nucleus, DNA Probes, Guanine, HeLa Cells, Humans, In Vitro Techniques, Oligonucleotide Probes, Poly A, RNA Polymerase II, RNA Processing, Post-Transcriptional, RNA, Messenger, Transcription, Genetic