rad51 polymerization reveals a new chromatin remodeling mechanismrad51聚合揭示了一个新的染色质重塑机制.pdfVIP

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rad51 polymerization reveals a new chromatin remodeling mechanismrad51聚合揭示了一个新的染色质重塑机制.pdf

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rad51 polymerization reveals a new chromatin remodeling mechanismrad51聚合揭示了一个新的染色质重塑机制

Rad51 Polymerization Reveals a New Chromatin Remodeling Mechanism Pauline Dupaigne1,2, Christophe Lavelle1,2, Anthony Justome1,2, Sophie Lafosse1,2, Gilles Mirambeau1,3, 1,2 ´ 1,2. 1,2. Marc Lipinski , Olivier Pietrement *, Eric Le Cam * ´ ´ ´ 1 CNRS, Laboratoire de Microscopie Moleculaire et Cellulaire, UMR 8126 Interactions Moleculaires et Cancer, Institut de cancerologie Gustave Roussy, Villejuif, France, 2 Univ. Paris-Sud, Villejuif, France, 3 Division de Biochimie, UFR des Sciences de la Vie, Univ. Pierre et Marie Curie, Paris, France Abstract Rad51 protein is a well known protagonist of homologous recombination in eukaryotic cells. Rad51 polymerization on single-stranded DNA and its role in presynaptic filament formation have been extensively documented. Rad51 polymerizes also on double-stranded DNA but the significance of this filament formation remains unclear. We explored the behavior of Saccharomyces cerevisiae Rad51 on dsDNA and the influence of nucleosomes on Rad51 polymerization mechanism to investigate its putative role in chromatin accessibility to recombination machinery. We combined biochemical approaches, transmission electron microscopy (TEM) and atomic force microscopy (AFM) for analysis of the effects of the Rad51 filament on chromatinized templates. Quantitative analyses clearly demonstrated the occurrence of chromatin remodeling during nucleoprotein filament formation. During Rad51 polymerization, recombinase proteins moved all the nucleosomal arrays in front of the progressing filament. This polymerization process had a powerful remodeling effect, as Rad51 destabilized the nucleosomes along consider