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Gijs Wuite

Gijs Wuite

Vrije Universiteit Amsterdam, The Netherlands

Title: Sliding sleeves of XRCC4–XLF bridge DNA and connect fragments of broken DNA

Biography

Biography: Gijs Wuite

Abstract

Non-homologous end joining (NHEJ) is the primary pathway for repairing DNA double-strand breaks (DSBs) in mammalian cells1 . Such breaks are formed, for example, during gene-segment rearrangements in the adaptive immune system or by cancer therapeutic agents. Although the core components of the NHEJ machinery are known, it has remained difficult to assess the specific roles of these components and the dynamics of bringing and holding the fragments of broken DNA together. The structurally similar XRCC4 and XLF proteins are proposed to assemble as highly dynamic filaments at (or near) DSBs2 . Here we show, using dualand quadruple-trap optical tweezers combined with fluorescence microscopy, how human XRCC4, XLF and XRCC4–XLF complexes interact with DNA in real time. We find that XLF stimulates the binding of XRCC4 to DNA, forming heteromeric complexes that diffuse swiftly along the DNA. Moreover, we find that XRCC4–XLF complexes robustly bridge two independent DNA molecules and that these bridges are able to slide along the DNA. These observations suggest that XRCC4–XLF complexes form mobile sleeve-like structures around DNA that can reconnect the broken ends very rapidly and hold them together. Understanding the dynamics and regulation of this mechanism will lead to clarification of how NHEJ proteins are involved in generating chromosomal translocations (Brouwer et al., Nature, doi:10.1038/nature18643 , 2016)  

References:

  1. Ineke Brouwer, Gerrit Sitters, Andrea Candelli, … , Mauro Modesti, Erwin J.G. Peterman, Gijs J.L. Wuite (2016) Sliding sleeves of XRCC4-XLF bridge DNA and connect fragments of broken DNA NATURE Volume:535 Issue: 7613
  2. Onno D. Broekmans, Graeme A. King, Greg J. Stephens, and Gijs J.L. Wuite (2016) DNA twist stability changes with magnesium(2+) concentration PHYS. REV. LETT.  Volume: 116   Issue: 25
  3. King, Graeme A; Peterman, Erwin J G; Wuite, Gijs J L (2016) Unravelling the structural plasticity of stretched DNA under torsional constraint NATURE COMMUNICATIONS Volume: 7 Pages: 11810
  4. Biebricher, Andreas S.; Heller, Iddo; … Peterman, Erwin J. G.; Wuite, Gijs J. L. (2015) The impact of DNA intercalators on DNA and DNA-processing enzymes elucidated through force-dependent binding kinetics NATURE COMMUNICATIONS Volume: 6 Article: 7304
  5. I Heller, G Sitters, OD Broekmans, G Farge, C Menges, W Wende, SW Hell, EJG Peterman & GJL Wuite. (2013) STED nanoscopy combined with optical tweezers reveals protein dynamics on densely covered DNA Nature Methods, 10.1038/nmeth.2599