Print Email Facebook Twitter Molecular mechanism of sequence-dependent stability of RecA filament Title Molecular mechanism of sequence-dependent stability of RecA filament Author Kim, S.H. Joo, C. Ha, T. Kim, D. Faculty Applied Sciences Department BN/Bionanoscience Date 2013-06-26 Abstract A is a DNA-dependent ATPase and mediates homologous recombination by first forming a filament on a single-stranded (ss) DNA. RecA binds preferentially to TGG repeat sequence, which resembles the recombination hot spot Chi (5'-GCTGGTGG-3') and is the most frequent pattern (GTG) of the codon usage in Escherichia coli. Because of the highly dynamic nature of RecA filament formation, which consists of filament nucleation, growth and shrinkage, we need experimental approaches that can resolve each of these processes separately to gain detailed insights into the molecular mechanism of sequence preference. By using a single-molecule fluorescence assay, we examined the effect of sequence on individual stages of nucleation, monomer binding and dissociation. We found that RecA does not recognize the Chi sequence as a nucleation site. In contrast, we observed that it is the reduced monomer dissociation that mainly determines the high filament stability on TGG repeats. This sequence dependence of monomer dissociation is well-correlated with that of ATP hydrolysis, suggesting that DNA sequence dictates filament stability through modulation of ATP hydrolysis. To reference this document use: http://resolver.tudelft.nl/uuid:ac2cebe3-1042-4fe0-a886-8c6eac741f97 DOI https://doi.org/10.1093/nar/gkt570 Publisher Oxford University Press ISSN 0305-1048 Source Nucleic Acids Research, 2013 Part of collection Institutional Repository Document type journal article Rights (c) 2013 The Author(s)This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Files PDF Joo_2013.pdf 740.32 KB Close viewer /islandora/object/uuid:ac2cebe3-1042-4fe0-a886-8c6eac741f97/datastream/OBJ/view