Template strand switching by T7 RNA polymerase.

The Journal of Biological Chemistry
M RongW T McAllister

Abstract

T7 RNA polymerase (RNAP) is able to traverse a variety of discontinuities in the template (T) strand of duplex DNA, including nicks, gaps, and branched junctions in which the 3' end of the T strand is not complementary to the non-template (NT) strand. The products represent a faithful copy of the T strand, with no insertions or deletions. On double-stranded templates having protruding 3' ends the polymerase is able to insert the free 3' end of the NT strand and to utilize this as a new T strand ("turn around transcription"), resulting in the anomalous production of high molecular weight transcripts. The capacity of T7 RNAP to bypass interruptions in the T strand depends upon the stability of the elongation complex. Sequences that are expected to stabilize a local RNA:DNA hybrid (such as the presence of a C6 tract in the T strand) dramatically reduce dissociation of the RNAP while still allowing the enzyme to insert a new 3' end. Similar effects on RNAP release are observed when the enzyme reaches the end of a template (i.e. when synthesizing runoff products), resulting in markedly different yields of RNA product during multiple rounds of transcription.

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Citations

Aug 10, 2004·Nature Biotechnology·Jijumon Chelliserrykattil, Andrew D Ellington
Jan 28, 2012·Nature Protocols·Pattabhiraman ShankaranarayananHinrich Gronemeyer
Nov 20, 2002·Journal of Virology·R C L OlsthoornJ J Bujarski
Jun 8, 2007·Proceedings of the National Academy of Sciences of the United States of America·Yi ZhouCraig T Martin
May 10, 2006·PLoS Computational Biology·Marc Bailly-BechetMassimo Vergassola
Aug 22, 2007·PLoS Computational Biology·Duncan P BrownKimmen Sjölander
Oct 17, 2007·PLoS Computational Biology·Jakob H HavgaardJan Gorodkin
Dec 21, 2010·Chemistry & Biology·Ryan C Todd, Stephen J Lippard
Jun 20, 2017·PLoS Genetics·Karin MerkKlaus Förstemann
May 3, 2018·Nucleic Acids Research·Samuel W SchaffterElisa Franco
Nov 19, 2019·Frontiers in Microbiology·Xueling LuBin Zhu
Sep 18, 2003·Journal of Cell Science·Sun-Kyung Lee, Peter J Hollenbeck
Aug 17, 2021·Journal of the American Chemical Society·Ze YuMichael Famulok

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