Artificial kinetochore beads establish a biorientation-like state in the spindle

Science. 2024 Sep 20;385(6715):1366-1375. doi: 10.1126/science.adn5428. Epub 2024 Sep 19.

Abstract

Faithful chromosome segregation requires biorientation, where the pair of kinetochores on the chromosome establish bipolar microtubule attachment. The integrity of the kinetochore, a macromolecular complex built on centromeric DNA, is required for biorientation, but components sufficient for biorientation remain unknown. Here, we show that tethering the outer kinetochore heterodimer NDC80-NUF2 to the surface of apolar microbeads establishes their biorientation-like state in mouse cells. NDC80-NUF2 microbeads align at the spindle equator and self-correct alignment errors. The alignment is associated with stable bipolar microtubule attachment and is independent of the outer kinetochore proteins SPC24-SPC25, KNL1, the Mis12 complex, inner kinetochore proteins, and Aurora. Larger microbeads align more rapidly, suggesting a size-dependent biorientation mechanism. This study demonstrates a biohybrid kinetochore design for synthetic biorientation of microscale particles in cells.

MeSH terms

  • Animals
  • Cell Cycle Proteins* / metabolism
  • Chromosome Segregation*
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism
  • Kinetochores* / metabolism
  • Mice
  • Microspheres*
  • Microtubule-Associated Proteins* / genetics
  • Microtubule-Associated Proteins* / metabolism
  • Microtubules* / metabolism
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Spindle Apparatus* / metabolism

Substances

  • Cell Cycle Proteins
  • Cytoskeletal Proteins
  • Microtubule-Associated Proteins
  • Nuclear Proteins
  • Hec1 protein, mouse
  • Nuf2 protein, mouse