CENP-C is a blueprint for constitutive centromere-associated network assembly within human kinetochores.
Bottom Line: A 16-subunit complex named the constitutive centromere-associated network (CCAN) creates the centromere-kinetochore interface.We identified crucial determinants of this interaction whose mutation prevented kinetochore localization of CENP-HIKM and of CENP-TW, another CCAN subcomplex.When considered together with previous observations, our data point to CENP-C as a blueprint for kinetochore assembly.
Affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.Show MeSH
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Mentions: In Fig. 1 (D and E), we demonstrated that kinetochore localization of CENP-TW depends on CENP-C. Kinetochore localization of CENP-TW had been previously shown to depend also on the CENP-HIKM complex (Hori et al., 2008; Basilico et al., 2014). Thus, we analyzed how CENP-C mutants impairing CENP-HIKM localization to kinetochores impacted kinetochore localization of CENP-TW. As for CENP-HK, wild-type CENP-C fully rescued kinetochore localization of CENP-TW, but there was a progressive reduction of CENP-TW kinetochore levels when CENP-C mutants were expressed (Fig. 4, A and B). In agreement with these observations, we readily identified CENP-TW in precipitates of GFP–CENP-C1–544-wt but not in precipitates of GFP–CENP-C1–544-4A (Fig. 3 C).
Affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.