Whole-proteome genetic analysis of dependencies in assembly of a vertebrate kinetochore

Abstract

Kinetochores orchestrate mitotic chromosome segregation. Here, we use quantitative mass spectrometry of mitotic chromosomes isolated from a comprehensive set of chicken DT40 mutants to examine the dependencies of 93 confirmed and putative kinetochore proteins for stable association with chromosomes. Clustering and network analysis reveal both known and unexpected aspects of coordinated behavior for members of kinetochore protein complexes. Surprisingly, CENP-T depends on CENP-N for chromosome localization. The Ndc80 complex exhibits robust correlations with all other complexes in a "core" kinetochore network. Ndc80 associated with CENP-T interacts with a cohort of Rod, zw10, and zwilch (RZZ)-interacting proteins that includes Spindly, Mad1, and CENP-E. This complex may coordinate microtubule binding with checkpoint signaling. Ndc80 associated with CENP-C forms the KMN (Knl1, Mis12, Ndc80) network and may be the microtubule-binding "workhorse" of the kinetochore. Our data also suggest that CENP-O and CENP-R may regulate the size of the inner kinetochore without influencing the assembly of the outer kinetochore.

DOI: 10.1083/jcb.201508072

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@inproceedings{Samejima2015WholeproteomeGA, title={Whole-proteome genetic analysis of dependencies in assembly of a vertebrate kinetochore}, author={Itaru Samejima and Christos Spanos and Fl{\'a}via de Lima Alves and Tetsuya Hori and Marinela Perpelescu and Juan Zou and Juri Rappsilber and Tatsuo Fukagawa and William C Earnshaw}, booktitle={The Journal of cell biology}, year={2015} }