Twisting and untwisting a single DNA molecule covered by RecA protein

扭转和解开 RecA 蛋白覆盖的单个 DNA 分子

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作者:Renaud Fulconis, Aurélien Bancaud, Jean-Francois Allemand, Vincent Croquette, Marie Dutreix, Jean-Louis Viovy

Abstract

We study dsDNA-RecA interactions by exerting forces in the pN range on single DNA molecules while the interstrand topological state is controlled owing to a magnetic tweezers setup. We show that unwinding a duplex DNA molecule induces RecA polymerization even at moderate force. Once initial polymerization has nucleated, the extent of RecA coverage still depends on the degree of supercoiling: exerting a positive or negative torsional constraint on the fiber forces partial depolymerization, with a strikingly greater stability when ATPgammaS is used as a cofactor instead of ATP. This nucleofilament's sensitivity to topology might be a way for the bacterial cell to limit consumption of precious RecA monomers when DNA damage is addressed through homologous recombination repair.

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