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Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
Published on: May 3, 2014
Multiple transcript cleavage precedes polymerase release in termination by RNA polymerase II
1Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom.
Cell
|June 8, 2001
Summary
RNA polymerase II (pol II) transcription termination involves two distinct events: pretermination cleavage (PTC) mediated by downstream sequences and polymerase disengagement requiring both PTC and a poly(A) site.
Area of Science:
- Molecular Biology
- Gene Expression
- Biochemistry
Background:
- Polyadenylation (poly(A)) signals are known to be essential for RNA polymerase II (pol II) transcription termination.
- The precise mechanisms governing pol II transcription termination remain largely unelucidated.
Purpose of the Study:
- To investigate the sequential events involved in RNA polymerase II transcription termination.
- To elucidate the roles of downstream sequences and poly(A) sites in the termination process.
Main Methods:
- Analysis of nascent transcripts from human beta- and epsilon-globin genes.
- Identification and characterization of pretermination cleavage (PTC) events.
Main Results:
- Pol II transcription termination comprises two distinct, temporally separated events.
- Pretermination cleavage (PTC) is facilitated by sequence tracts downstream of the poly(A) site, causing heterogeneous transcript cleavage.
- Polymerase disengagement from the DNA template necessitates prior transcription of both a PTC sequence and a functional poly(A) site.
Conclusions:
- Transcription termination by RNA polymerase II is a multi-step process.
- Sequence elements downstream of the poly(A) site play a critical role in initiating termination through pretermination cleavage.
- Efficient termination requires the coordinated action of both pretermination cleavage and polyadenylation signals.
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