Local control of intracellular microtubule dynamics by EB1 photodissociation

Nat Cell Biol. 2018 Mar;20(3):252-261. doi: 10.1038/s41556-017-0028-5. Epub 2018 Jan 29.

Abstract

End-binding proteins (EBs) are adaptors that recruit functionally diverse microtubule plus-end-tracking proteins (+TIPs) to growing microtubule plus ends. To test with high spatial and temporal accuracy how, when and where +TIP complexes contribute to dynamic cell biology, we developed a photo-inactivated EB1 variant (π-EB1) by inserting a blue-light-sensitive protein-protein interaction module between the microtubule-binding and +TIP-binding domains of EB1. π-EB1 replaces endogenous EB1 function in the absence of blue light. By contrast, blue-light-mediated π-EB1 photodissociation results in rapid +TIP complex disassembly, and acutely and reversibly attenuates microtubule growth independent of microtubule end association of the microtubule polymerase CKAP5 (also known as ch-TOG and XMAP215). Local π-EB1 photodissociation allows subcellular control of microtubule dynamics at the second and micrometre scale, and elicits aversive turning of migrating cancer cells. Importantly, light-mediated domain splitting can serve as a template to optically control other intracellular protein activities.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Carcinoma, Non-Small-Cell Lung / genetics
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • Carcinoma, Non-Small-Cell Lung / pathology
  • Cell Line, Tumor
  • Cell Movement*
  • Humans
  • Lung Neoplasms / genetics
  • Lung Neoplasms / metabolism*
  • Lung Neoplasms / pathology
  • Microscopy, Fluorescence
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubule-Associated Proteins / radiation effects
  • Microtubules / genetics
  • Microtubules / metabolism*
  • Microtubules / pathology
  • Microtubules / radiation effects
  • Optogenetics*
  • Photolysis
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Signal Transduction
  • Time Factors
  • Time-Lapse Imaging

Substances

  • CKAP5 protein, human
  • MAPRE1 protein, human
  • Microtubule-Associated Proteins