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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
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Eukaryotic Rad50 functions as a rod-shaped dimer.

Young Bong Park1, Marcel Hohl2, Michał Padjasek3

  • 1Department of Life Sciences, Pohang University of Science and Technology, Pohang, South Korea.

Nature Structural & Molecular Biology
|January 31, 2017
PubMed
Summary
This summary is machine-generated.

The Rad50 hook and coiled-coil interfaces promote intracomplex assembly of the Mre11 complex. This intracomplex structure is the functional form, essential for Tel1 activation and DNA repair.

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Area of Science:

  • Molecular biology
  • Structural biology
  • Biochemistry

Background:

  • The Rad50 hook interface is vital for the Mre11 complex's assembly and function.
  • Previous studies proposed both intracomplex and intercomplex interactions for Rad50 molecules.

Purpose of the Study:

  • To determine the structure of the human Rad50 hook and coiled-coil domains.
  • To elucidate the predominant assembly state and functional implications of Rad50 interactions within the Mre11 complex.

Main Methods:

  • X-ray crystallography to determine the structure of human Rad50 hook and coiled-coil domains.
  • Functional assays in yeast to assess the impact of interface disruption on Tel1 activation and DNA repair.

Main Results:

  • The predominant structure identified is intracomplex, with parallel coiled coils forming a rod.
  • A novel interface within the coiled-coil domains stabilizes Rad50 protomer interactions in the dimeric assembly.
  • Disrupting the coiled-coil interface in yeast impaired Tel1 activation but not DNA repair.

Conclusions:

  • The hook and coiled-coil interfaces work together to promote intracomplex assembly.
  • The intracomplex form is identified as the functional state of the Mre11 complex.
  • Coordinated action of these interfaces is critical for Mre11 complex function, including Tel1 activation.