The highly processive DNA polymerase of bacteriophage T5. Role of the unique N and C termini

The Journal of Biological Chemistry
Nathalie AndraosC C Richardson

Abstract

The DNA polymerase encoded by bacteriophage T5 has been reported previously to be processive and to catalyze extensive strand displacement synthesis. The enzyme, purified from phage-infected cells, did not require accessory proteins for these activities. Although T5 DNA polymerase shares extensive sequence homology with Escherichia coli DNA polymerase I and T7 DNA polymerase, it contains unique regions of 130 and 71 residues at its N and C termini, respectively. We cloned the gene encoding wild-type T5 DNA polymerase and characterized the overproduced protein. We also examined the effect of N- and C-terminal deletions on processivity and strand displacement synthesis. T5 DNA polymerase lacking its N-terminal 30 residues resembled the wild-type enzyme albeit with a 2-fold reduction in polymerase activity. Deletion of 24 residues at the C terminus resulted in a 30-fold reduction in polymerase activity on primed circular DNA, had dramatically reduced processivity, and was unable to carry out strand displacement synthesis. Deletion of 63 residues at the C terminus resulted in a 20,000-fold reduction in polymerase activity. The 3' to 5' double-stranded DNA exonuclease activity associated with T5 DNA polymerase was reduced by a facto...Continue Reading

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Citations

Jun 24, 2011·The Journal of Biological Chemistry·Barak AkabayovCharles C Richardson
Jul 12, 2011·Nucleic Acids Research·Darius Kazlauskas, Ceslovas Venclovas
Feb 26, 2013·Nucleic Acids Research·Io Nam WongCyril M Sanders
Mar 21, 2009·Annual Review of Biochemistry·Samir M Hamdan, Charles C Richardson
Jun 8, 2012·PloS One·Michael J MoserThomas W Schoenfeld
Jun 4, 2015·Annual Review of Biochemistry·Charles C Richardson
Nov 20, 2013·FEBS Open Bio·Olga PiskarevaVadim Schmatchenko
Jun 30, 2006·The Journal of Biological Chemistry·Seung-Joo LeeCharles C Richardson
Jul 18, 2021·Science Advances·Xiao YiMichael Travisano
Nov 10, 2009·Trends in Microbiology·Thomas SchoenfeldDavid Mead
Sep 3, 2010·Mitochondrion·Katrin ViikovJuhan Sedman

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