Aurora B switches relative strength of kinetochore–microtubule attachment modes for error correction

Author:

Doodhi Harinath1ORCID,Kasciukovic Taciana1ORCID,Clayton Lesley1ORCID,Tanaka Tomoyuki U.1ORCID

Affiliation:

1. Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee, UK

Abstract

To establish chromosome biorientation, aberrant kinetochore–microtubule interaction must be resolved (error correction) by Aurora B kinase. Aurora B differentially regulates kinetochore attachment to the microtubule plus end and its lateral side (end-on and lateral attachment, respectively). However, it is still unclear how kinetochore–microtubule interactions are exchanged during error correction. Here, we reconstituted the budding yeast kinetochore–microtubule interface in vitro by attaching the Ndc80 complexes to nanobeads. These Ndc80C nanobeads recapitulated in vitro the lateral and end-on attachments of authentic kinetochores on dynamic microtubules loaded with the Dam1 complex. This in vitro assay enabled the direct comparison of lateral and end-on attachment strength and showed that Dam1 phosphorylation by Aurora B makes the end-on attachment weaker than the lateral attachment. Similar reconstitutions with purified kinetochore particles were used for comparison. We suggest the Dam1 phosphorylation weakens interaction with the Ndc80 complex, disrupts the end-on attachment, and promotes the exchange to a new lateral attachment, leading to error correction.

Funder

Wellcome Trust

Medical Research Council

Publisher

Rockefeller University Press

Subject

Cell Biology

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