Print Email Facebook Twitter RecA filament maintains structural integrity using ATP-driven internal dynamics Title RecA filament maintains structural integrity using ATP-driven internal dynamics Author Kim, S.H. (TU Delft BN/Cees Dekker Lab; Sogang University; Kavli institute of nanoscience Delft) Ahn, Tak Kyoon (Sogang University) Cui, T.J. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft) Chauhan, S. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft) Sung, Jaeyoung (Chung-Ang University) Joo, C. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft) Kim, Doseok (Sogang University) Date 2017-09-01 Abstract At the core of homologous DNA repair, RecA catalyzes the strand exchange reaction. This process is initiated by a RecA loading protein, which nucleates clusters of RecA proteins on single-stranded DNA. Each cluster grows to cover the single-stranded DNA but may leave 1- to 2-nucleotide (nt) gaps between the clusters due to three different structural phases of the nucleoprotein filaments. It remains to be revealed how RecA proteins eliminate the gaps to make a seamless kilobase-long filament. We develop a single-molecule fluorescence assay to observe the novel internal dynamics of the RecA filament. We directly observe the structural phases of individual RecA filaments and find that RecA proteins move their positions along the substrate DNA to change the phase of the filament. This reorganization process, which is a prerequisite step for interjoining of two adjacent clusters, requires adenosine triphosphate hydrolysis and is tightly regulated by the recombination hotspot, Chi. Furthermore, RecA proteins recognize and self-align to a 3-nt-period sequence pattern of TGG. This sequence-dependent phase bias may help the RecA filament to maintain structural integrity within the kilobase-long filament for accurate homology search and strand exchange reaction. To reference this document use: http://resolver.tudelft.nl/uuid:437fce8a-fd3c-4513-b442-7bd40aeda113 DOI https://doi.org/10.1126/sciadv.1700676 ISSN 2375-2548 Source Science Advances, 3 (9) Part of collection Institutional Repository Document type journal article Rights © 2017 S.H. Kim, Tak Kyoon Ahn, T.J. Cui, S. Chauhan, Jaeyoung Sung, C. Joo, Doseok Kim Files PDF e1700676.full.pdf 2.18 MB Close viewer /islandora/object/uuid:437fce8a-fd3c-4513-b442-7bd40aeda113/datastream/OBJ/view