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• W2050876417 endingPage "1386" @default.
• W2050876417 startingPage "1379" @default.
• W2050876417 abstract "Phage Phi29 encodes a DNA-dependent DNA polymerase belonging to the eukaryotic-type (family B) subgroup of DNA polymerases that use a protein as the primer for initiation of DNA synthesis. In one of the most important motifs present in the 3'-->5' exonucleolytic domain of proofreading DNA polymerases, the ExoII motif, Phi29 DNA polymerase contains three amino acid residues, Y59, H61 and F69, which are highly conserved among most proofreading DNA polymerases. These residues have recently been shown to be involved in proper stabilization of the primer terminus at the 3'-->5' exonuclease active site. Here we investigate by means of site-directed mutagenesis the role of these three residues in reactions that are specific for DNA polymerases utilizing a protein-primed DNA replication mechanism. Mutations introduced at residues Y59, H61 and F69 severely affected the protein-primed replication capacity of Phi29 DNA polymerase. For four of the mutants, namely Y59L, H61L, H61R and F69S, interaction with the terminal protein was affected, leading to few initiation and transition products. These findings, together with the specific conservation of Y59, H61 and F69 among DNA polymerases belonging to the protein-primed subgroup, strongly suggest a functional role of these amino acid residues in the DNA polymerase-terminal protein interaction." @default.
• W2050876417 created "2016-06-24" @default.
• W2050876417 creator A5020258455 @default.
• W2050876417 creator A5046841135 @default.
• W2050876417 creator A5089344832 @default.
• W2050876417 creator A5091770895 @default.
• W2050876417 date "2002-03-15" @default.
• W2050876417 modified "2023-10-01" @default.
• W2050876417 title "Phi29 DNA polymerase residues Tyr59, His61 and Phe69 of the highly conserved ExoII motif are essential for interaction with the terminal protein" @default.
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• W2050876417 doi "https://doi.org/10.1093/nar/30.6.1379" @default.
• W2050876417 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/101362" @default.
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• W2050876417 hasPublicationYear "2002" @default.
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