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Each human somatic cell contains 6 billion base pairs of DNA. Each base pair is 0.34 nm long, meaning each diploid cell contains a staggering 2 meters of DNA. This long DNA strand is packed inside a nucleus measuring only 10-20 microns in diameter with the help of specialized DNA-binding proteins called histones. Together they form a compact DNA-protein complex called chromatin. The chromatin is further compacted into higher-order structures. The highest level of compaction is achieved during...
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Each human somatic cell contains 6 billion base-pairs of DNA. Each base-pair is 0.34 nm long, which means that each diploid cell contains a staggering 2 meters of DNA. How is such a long DNA strand packed inside a nucleus measuring only 10 - 20 microns in diameter? 
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Studying DNA Looping by Single-Molecule FRET
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DNA loop extrusion by human cohesin.

Iain F Davidson1, Benedikt Bauer1, Daniela Goetz1

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Summary
This summary is machine-generated.

Human cohesin complexes actively extrude DNA into loops, forming crucial chromatin structures. This process, dependent on cohesin

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • Eukaryotic genomes fold into loops and topologically associating domains, influencing gene regulation and recombination.
  • These structures rely on cohesin, a DNA-entrapping ATPase complex, proposed to form loops via extrusion.
  • While condensin exhibits loop extrusion in mitosis, cohesin's activity remained unconfirmed.

Purpose of the Study:

  • To investigate whether cohesin can form DNA loops through extrusion.
  • To elucidate the mechanism and requirements for cohesin-mediated loop formation.

Main Methods:

  • Biochemical reconstitution of human cohesin complexes.
  • Real-time observation of DNA loop formation by single cohesin complexes.
  • Assays to determine dependence on ATPase activity, NIPBL-MAU2, and topological entrapment.

Main Results:

  • Single human cohesin complexes extrude DNA loops at rates up to 2.1 kilobase pairs per second.
  • Loop formation and maintenance require cohesin's ATPase activity and the NIPBL-MAU2 factor.
  • Topological entrapment of DNA by cohesin is not necessary for loop extrusion.
  • Cohesin and NIPBL-MAU2 localize at the base of extruded loops, confirming their role in extrusion.

Conclusions:

  • Cohesin and NIPBL-MAU2 form an active holoenzyme capable of extruding DNA into loops during interphase.
  • This loop extrusion mechanism, independent of topological entrapment, is fundamental to chromatin organization.