Nucleotide selection by the Y-family DNA polymerase Dpo4 involves template translocation and misalignment.

ABSTRACT: Y-family DNA polymerases play a crucial role in translesion DNA synthesis. Here, we have characterized the binding kinetics and conformational dynamics of the Y-family polymerase Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) using single-molecule fluorescence. We find that in the absence of dNTPs, the binary complex shuttles between two different conformations within ∼1 s. These data are consistent with prior crystal structures in which the nucleotide binding site is either occupied by the terminal base pair (preinsertion conformation) or empty following Dpo4 translocation by 1 base pair (insertion conformation). Most interestingly, on dNTP binding, only the insertion conformation is observed and the correct dNTP stabilizes this complex compared with the binary complex, whereas incor

SUBMITTER: Brenlla A 

PROVIDER: S-EPMC3936744 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature