Real-time Observation of Strand Exchange Reaction with High Spatiotemporal Resolution

Overview

Journal Structure

Publisher Cell Press

Specialties Biochemistry
Biotechnology
Cell Biology
Molecular Biology

Date 2011 Aug 11

PMID 21827943

Citations 36

Authors

Kaushik Ragunathan

Chirlmin Joo

Taekjip Ha

Affiliations

Soon will be listed here.

Abstract

RecA binds to single-stranded (ss) DNA to form a helical filament that catalyzes strand exchange with a homologous double-stranded (ds) DNA. The study of strand exchange in ensemble assays is limited by the diffusion limited homology search process, which masks the subsequent strand exchange reaction. We developed a single-molecule fluorescence assay with a few base-pair and millisecond resolution that can separate initial docking from the subsequent propagation of joint molecule formation. Our data suggest that propagation occurs in 3 bp increments with destabilization of the incoming dsDNA and concomitant pairing with the reference ssDNA. Unexpectedly, we discovered the formation of a dynamic complex between RecA and the displaced DNA that remains bound transiently after joint molecule formation. This finding could have important implications for the irreversibility of strand exchange. Our model for strand exchange links structural models of RecA to its catalytic function.

Citing Articles

Homology recognition without double-stranded DNA-strand separation in D-loop formation by RecA.

Shibata T, Ikawa S, Iwasaki W, Sasanuma H, Masai H, Hirota K Nucleic Acids Res. 2024; 52(5):2565-2577.

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Flanking strand separation activity of RecA nucleoprotein filaments in DNA strand exchange reactions.

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