RAMA1 is a novel kinetochore protein involved in kinetochore-microtubule attachment

J Cell Sci. 2009 Jul 15;122(Pt 14):2436-45. doi: 10.1242/jcs.051912. Epub 2009 Jun 23.

Abstract

During mitosis, kinetochores need to attach to microtubules emanating from spindle poles. Several protein complexes have been shown to mediate the kinetochore-microtubule interaction. However, with the continually growing number of newly identified kinetochore proteins, it is unclear whether all major components of the kinetochore-microtubule interface have been identified. We therefore performed a high-throughput RNAi screen to identify additional factors involved in kinetochore-microtubule attachment, and identified RAMA1 as a novel regulator of this process. Depletion of RAMA1 results in severe chromosome alignment defects and a checkpoint-dependent mitotic arrest. We show that this is due to reduced kinetochore-microtubule attachments. RAMA1 localizes to the spindle and to outer kinetochores throughout all phases of mitosis and is recruited to kinetochores by the core kinetochore-microtubule attachment factor Hec1. Interestingly, unlike Hec1, the association of RAMA1 with kinetochores is highly dynamic, suggesting that it is not a structural component of the kinetochore. Consistent with this, all other kinetochore proteins tested do not require RAMA1 for their kinetochore localization. Taken together, these results identify RAMA1 as a novel kinetochore protein and suggest that RAMA1 may have a direct role in mediating kinetochore-microtubule interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins
  • Chromosome Segregation
  • Cytoskeletal Proteins
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism*
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / metabolism*
  • Mitosis / physiology*
  • Nuclear Proteins / metabolism
  • RNA Interference
  • Recombinant Fusion Proteins / metabolism
  • Spindle Apparatus / metabolism
  • Time Factors
  • Transfection

Substances

  • Cell Cycle Proteins
  • Cytoskeletal Proteins
  • Microtubule-Associated Proteins
  • NDC80 protein, human
  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Ska3 protein, human