Synergistic Coordination of Chromatin Torsional Mechanics and Topoisomerase Activity

Cell. 2019 Oct 17;179(3):619-631.e15. doi: 10.1016/j.cell.2019.09.034.

Abstract

DNA replication in eukaryotes generates DNA supercoiling, which may intertwine (braid) daughter chromatin fibers to form precatenanes, posing topological challenges during chromosome segregation. The mechanisms that limit precatenane formation remain unclear. By making direct torque measurements, we demonstrate that the intrinsic mechanical properties of chromatin play a fundamental role in dictating precatenane formation and regulating chromatin topology. Whereas a single chromatin fiber is torsionally soft, a braided fiber is torsionally stiff, indicating that supercoiling on chromatin substrates is preferentially directed in front of the fork during replication. We further show that topoisomerase II relaxation displays a strong preference for a single chromatin fiber over a braided fiber. These results suggest a synergistic coordination-the mechanical properties of chromatin inherently suppress precatenane formation during replication elongation by driving DNA supercoiling ahead of the fork, where supercoiling is more efficiently removed by topoisomerase II. VIDEO ABSTRACT.

Keywords: DNA supercoiling; angular optical trap; braiding; chromatin; fork rotation; magnetic tweezers; replication; topoisomerase; torque; torsional modulus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / chemistry*
  • Chromatin / metabolism
  • DNA Replication
  • DNA Topoisomerases, Type II / metabolism*
  • DNA, Superhelical / chemistry
  • HeLa Cells
  • Humans
  • Optical Tweezers
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Torque*

Substances

  • Chromatin
  • DNA, Superhelical
  • Saccharomyces cerevisiae Proteins
  • DNA Topoisomerases, Type II