Single-molecule investigation of substrate binding kinetics and protein conformational dynamics of a B-family replicative DNA polymerase.

ABSTRACT: Replicative DNA polymerases use a complex, multistep mechanism for efficient and accurate DNA replication as uncovered by intense kinetic and structural studies. Recently, single-molecule fluorescence spectroscopy has provided new insights into real time conformational dynamics utilized by DNA polymerases during substrate binding and nucleotide incorporation. We have used single-molecule Förster resonance energy transfer techniques to investigate the kinetics and conformational dynamics of Sulfolobus solfataricus DNA polymerase B1 (PolB1) during DNA and nucleotide binding. Our experiments demonstrate that this replicative polymerase can bind to DNA in at least three conformations, corresponding to an open and closed conformation of the finger domain as well as a conformation with the DNA s

SUBMITTER: Maxwell BA 

PROVIDER: S-EPMC3630881 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature