Structures of phi29 DNA polymerase complexed with substrate: the mechanism of translocation in B-family polymerases
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 cryst...
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Published in | The EMBO journal Vol. 26; no. 14; pp. 3494 - 3505 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Chichester, UK
John Wiley & Sons, Ltd
25.07.2007
Blackwell Publishing Ltd Nature Publishing Group |
Subjects | |
Online Access | Get full text |
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Summary: | 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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Bibliography: | istex:C9F69990D20DC7B316034FC350271F89AAFE05E9 ArticleID:EMBJ7601780 Supplementary MovieSupplementary Figure S1Supplementary Figure S2Supplementary Figure S3Supplementary DataSupplementary Data ark:/67375/WNG-9X3J6B79-M ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 DE-AC02-98CH10886 BNL-80556-2008-JA Doe - Office Of Science Present address: Pfizer Inc., 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA |
ISSN: | 0261-4189 1460-2075 |
DOI: | 10.1038/sj.emboj.7601780 |