Abstract
The first DNA joint formed in homologous recombination processes is a D-loop. Saccharomyces cerevisiae RDH54/TID1-encoded product, a Swi2/Snf2-like factor involved in recombination, is shown here to promote D-loop formation with Rad51 recombinase. Physical interaction between Rdh54 and Rad51 is functionally important because Rdh54 does not enhance the recombinase activity of the Escherichia coli RecA protein. Robust dsDNA-activated ATPase activity in Rdh54 generates unconstrained negative and positive supercoils in DNA. Efficient D-loop formation occurs with even topologically relaxed DNA, suggesting that via specific protein-protein interactions, the negative supercoils produced by Rdh54 are used by Rad51 for making DNA joints.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Triphosphatases / metabolism
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DNA / metabolism
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DNA Helicases
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DNA Repair Enzymes
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DNA Topoisomerases
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DNA Topoisomerases, Type I / metabolism
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DNA, Superhelical / metabolism
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Electrophoresis, Agar Gel
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Electrophoresis, Gel, Two-Dimensional
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Escherichia coli / metabolism
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Gene Expression Regulation, Fungal
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Immunoblotting
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Plasmids / metabolism
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Protein Binding
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Rad51 Recombinase
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Rec A Recombinases / metabolism
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Recombination, Genetic
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins*
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Time Factors
Substances
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DNA, Superhelical
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DNA-Binding Proteins
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Fungal Proteins
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Saccharomyces cerevisiae Proteins
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DNA
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RAD51 protein, S cerevisiae
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Rad51 Recombinase
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Rec A Recombinases
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Adenosine Triphosphatases
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RAD54 protein, S cerevisiae
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DNA Helicases
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DNA Topoisomerases
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RDH54 protein, S cerevisiae
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DNA Topoisomerases, Type I
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DNA Repair Enzymes