RecA filament sliding on DNA facilitates homology search

Author:

Ragunathan Kaushik123,Liu Cheng4,Ha Taekjip12345

Affiliation:

1. Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, United States

2. Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, United States

3. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, United States

4. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, United States

5. Howard Hughes Medical Institute, University of Illinois at Urbana-Champaign, Urbana, United States

Abstract

During homologous recombination, RecA forms a helical filament on a single stranded (ss) DNA that searches for a homologous double stranded (ds) DNA and catalyzes the exchange of complementary base pairs to form a new heteroduplex. Using single molecule fluorescence imaging tools with high spatiotemporal resolution we characterized the encounter complex between the RecA filament and dsDNA. We present evidence in support of the ‘sliding model’ wherein a RecA filament diffuses along a dsDNA track. We further show that homology can be detected during sliding. Sliding occurs with a diffusion coefficient of approximately 8000 bp2/s allowing the filament to sample several hundred base pairs before dissociation. Modeling suggests that sliding can accelerate homology search by as much as 200 fold. Homology recognition can occur for as few as 6 nt of complementary basepairs with the recognition efficiency increasing for higher complementarity. Our data represents the first example of a DNA bound multi-protein complex which can slide along another DNA to facilitate target search.

Funder

National Science Foundation

National Institutes of Health

Howard Hughes Medical Institute

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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