Dynamic investigation of DNA bending and wrapping by type II topoisomerases

ORAL

Abstract

Type II topoisomerases catalyze DNA decatenation and unwinding which is crucial for cell division, and therefore type II topoisomerases are some of the main targets of anti-cancer drugs. A recent crystal structure shows that, during the catalytic cycle, a yeast type II topoimerase can bend a 10 base pair DNA segment by up to 150 degrees. Bacterial gyrase, another type II topoisomerase, can wrap DNA into a tight 180 degree turn. Bending a stiff polymer like DNA requires considerable energy and could represent the rate limiting step in the catalytic (topological) cycle. Using modified deoxyribonucleotides in PCR reactions, stiffer DNA fragments have been produced and used as substrates for topoisomerase II-mediated relaxation of plectonemes introduced in single molecules using magnetic tweezers. The wrapping ability of gyrase decreases for diamino-purine-substituted DNA in which every base pair has three hydrogen-bonds. The overall rate of relaxation of plectonemes by recombinant human topoisomerase II alpha also decreases. These results reveal the dynamic properties of DNA bending and wrapping by type II topisomerases and suggest that A:T base pair melting is a rate determining step for bending and wrapping.

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Authors

  • Qing Shao

    Physics Department, Emory University

  • Milind Purohit

    Univ of South Carolina, Benedict College, Sc 29204, Univ. South Carolina, Benedict College, Univ South Carolina, GA Tech, Central Microscopy Research Facility, University of Iowa, Iowa City, Iowa 52242, USA, Department of Chemistry, University of Sri Jayewardenepura, Nugegoda, Sri Lanka, Department of Physics and Astronomy, Georgia State University, Atlanta, Georgia 30303, USA, Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL 35294, Department of Physics, University of Alabama at Birmingham, Birmingham, AL 35294, College of William \& Mary, Harvard University, Benedict College, SC 29204, Massachusetts Institute of Technology, University of Applied Science, Bielefeld, Germany, Francis Marion University, Physics Dept., Emory University, Emory University, Formerly Emory University, currently UCLA, Physics Department, Georgia State University, Univ. of Georgia, Dept. of Chemistry, Universidad Nacional de Quilmes, Argentina, Pr, Dr, Derpartment of Physics, Florida A\&M University, Tallahassee, FL-32307, Department of Physics, Emory University, Georgia Institute of Technology, Naval Research Laboratory, University of Alabama, Tsinghua University, Lawrence Berkeley National Laboratory, Vanderbilt University, Jagellonian Univ., Univ. of Bonn, North Carolina A\&T State Univ., North Carolina Central Univ., Duke Univ. and TUNL, Georgia State University, Dept of Physics, Emory University, Cell Biology Department, Emory University, Physics Department, Emory University, University of South Carolina

  • Milind Purohit

    Univ of South Carolina, Benedict College, Sc 29204, Univ. South Carolina, Benedict College, Univ South Carolina, GA Tech, Central Microscopy Research Facility, University of Iowa, Iowa City, Iowa 52242, USA, Department of Chemistry, University of Sri Jayewardenepura, Nugegoda, Sri Lanka, Department of Physics and Astronomy, Georgia State University, Atlanta, Georgia 30303, USA, Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL 35294, Department of Physics, University of Alabama at Birmingham, Birmingham, AL 35294, College of William \& Mary, Harvard University, Benedict College, SC 29204, Massachusetts Institute of Technology, University of Applied Science, Bielefeld, Germany, Francis Marion University, Physics Dept., Emory University, Emory University, Formerly Emory University, currently UCLA, Physics Department, Georgia State University, Univ. of Georgia, Dept. of Chemistry, Universidad Nacional de Quilmes, Argentina, Pr, Dr, Derpartment of Physics, Florida A\&M University, Tallahassee, FL-32307, Department of Physics, Emory University, Georgia Institute of Technology, Naval Research Laboratory, University of Alabama, Tsinghua University, Lawrence Berkeley National Laboratory, Vanderbilt University, Jagellonian Univ., Univ. of Bonn, North Carolina A\&T State Univ., North Carolina Central Univ., Duke Univ. and TUNL, Georgia State University, Dept of Physics, Emory University, Cell Biology Department, Emory University, Physics Department, Emory University, University of South Carolina