Heterogeneity in E. coli RecBCD Helicase-DNA Binding and Base Pair Melting.

ABSTRACT: E. coli RecBCD, a helicase/nuclease involved in double stranded (ds) DNA break repair, binds to a dsDNA end and melts out several DNA base pairs (bp) using only its binding free energy. We examined RecBCD-DNA initiation complexes using thermodynamic and structural approaches. Measurements of enthalpy changes for RecBCD binding to DNA ends possessing pre-melted ssDNA tails of increasing length suggest that RecBCD interacts with ssDNA as long as 17-18 nucleotides and can melt at least 10-11 bp upon binding a blunt DNA end. Cryo-EM structures of RecBCD alone and in complex with a blunt-ended dsDNA show significant conformational heterogeneities associated with the RecB nuclease domain (RecB^Nuc) and the RecD subunit. In the absence of DNA, 56% of RecBCD molecules show no density for

SUBMITTER: Hao L 

PROVIDER: S-EPMC8996488 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature