TY - JOUR
T1 - Functional interdependence of DNA polymerizing and 3'→5' exonucleolytic activities in Pyrococcus furiosus DNA polymerase I
AU - Komori, Kayoko
AU - Ishino, Yoshizumi
PY - 2000
Y1 - 2000
N2 - Pyrococcus furiosus DNA polymerase I (Pol BI) belongs to the family B (α-like) DNA polymerases and has a strong 3'→5' exonucleolytic activity, in addition to its DNA polymerizing activity. To understand the relationship between the structure and function of this DNA polymerase, three deletion mutants, Δ1 (ΔLeu746-Ser775), Δ2 (ΔLeu717-Ser775) and Δ3 (ΔHis672-Ser775), and two substituted mutants of Asp405, D405A and D405E, were constructed. These substitutions affected both the DNA polymerizing and the 3'→5' exonucleolytic activities. The Δ1 mutant protein had DNA polymerizing activity with higher specific activity than that of the wild-type Pol BI, but retained only 10% of the exonucleolytic activity of the wild-type. The other two deletion mutants lost most of both activities. These results suggest that the DNA polymerizing and exonucleolytic activities are closely related to each other in the folded structure of this DNA polymerase, as proposed in the family B DNA polymerases.
AB - Pyrococcus furiosus DNA polymerase I (Pol BI) belongs to the family B (α-like) DNA polymerases and has a strong 3'→5' exonucleolytic activity, in addition to its DNA polymerizing activity. To understand the relationship between the structure and function of this DNA polymerase, three deletion mutants, Δ1 (ΔLeu746-Ser775), Δ2 (ΔLeu717-Ser775) and Δ3 (ΔHis672-Ser775), and two substituted mutants of Asp405, D405A and D405E, were constructed. These substitutions affected both the DNA polymerizing and the 3'→5' exonucleolytic activities. The Δ1 mutant protein had DNA polymerizing activity with higher specific activity than that of the wild-type Pol BI, but retained only 10% of the exonucleolytic activity of the wild-type. The other two deletion mutants lost most of both activities. These results suggest that the DNA polymerizing and exonucleolytic activities are closely related to each other in the folded structure of this DNA polymerase, as proposed in the family B DNA polymerases.
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U2 - 10.1093/protein/13.1.41
DO - 10.1093/protein/13.1.41
M3 - Article
C2 - 10679529
AN - SCOPUS:0033956483
SN - 0269-2139
VL - 13
SP - 41
EP - 47
JO - Protein Engineering
JF - Protein Engineering
IS - 1
ER -