Structures of Phi29 DNA Polymerase Complexed with Substrate: the Mechanism of Translocation in B-family Polymerases

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2007 Jul 6

PMID 17611604

Citations 94

Authors

Andrea J Berman

Satwik Kamtekar

Jessica L Goodman

Jose M Lazaro

Miguel de Vega

Luis Blanco

Margarita Salas

Thomas A Steitz

Affiliations

Soon will be listed here.

Abstract

Replicative DNA polymerases (DNAPs) move along template DNA in a processive manner. The structural basis of the mechanism of translocation has been better studied in the A-family of polymerases than in the B-family of replicative polymerases. To address this issue, we have determined the X-ray crystal structures of phi29 DNAP, a member of the protein-primed subgroup of the B-family of polymerases, complexed with primer-template DNA in the presence or absence of the incoming nucleoside triphosphate, the pre- and post-translocated states, respectively. Comparison of these structures reveals a mechanism of translocation that appears to be facilitated by the coordinated movement of two conserved tyrosine residues into the insertion site. This differs from the mechanism employed by the A-family polymerases, in which a conserved tyrosine moves into the templating and insertion sites during the translocation step. Polymerases from the two families also interact with downstream single-stranded template DNA in very different ways.

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