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Review
Subject Categories: RNA | Structural Biology
The EMBO Journal (2006) 25, 3458–3468, doi:10.1038/sj.emboj.7601211
Visualizing polynucleotide polymerase machines at work
Thomas A Steitz
Departments of Molecular Biophysics & Biochemistry and Chemistry, Yale University, and Howard Hughes Medical Institute, New Haven, CT, USA

To whom correspondence should be addressed
Thomas A Steitz, Departments of Molecular Biophysics & Biochemistry and Chemistry, Yale University, and Howard Hughes Medical Institute, New Haven, CT 06520-8114, USA. Tel.: +1 203 432 5617/5619; Fax: +1 203 432 3282; E-mail: eatherton@csb.yale.edu

Received 22 March 2006; Accepted 29 May 2006.
Abstract
The structures of T7 RNA polymerase (T7 RNAP) captured in the initiation and elongation phases of transcription, that of phi29 DNA polymerase bound to a primer protein and those of the multisubunit RNAPs bound to initiating factors provide insights into how these proteins can initiate RNA synthesis and synthesize 6–10 nucleotides while remaining bound to the site of initiation. Structural insight into the translocation of the product transcript and the separation of the downstream duplex DNA is provided by the structures of the four states of nucleotide incorporation. Single molecule and biochemical studies show a distribution of primer terminus positions that is altered by the binding of NTP and PPi ligands. This article reviews the insights that imaging the structure of polynucleotide polymerases at different steps of the polymerization reaction has provided on the mechanisms of the polymerization reaction. Movies are shown that allow the direct visualization of the conformational changes that the polymerases undergo during the different steps of polymerization.
Keywords: DNA polymerase, polymerase translocation, RNA polymerase
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