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. 2013 Jan 7;200(1):45–60. doi: 10.1083/jcb.201210106

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© 2013 Hori et al.

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Figure 7. — Model of kinetochore assembly in the engineered kinetochores. (A) Molecular architecture of the CENP-T or CENP-C N terminus–derived kinetochores. CENP-A and CCAN proteins are not detected in both the CENP-T or CENP-C N terminus–derived kinetochores. Function of these kinetochores is sensitive to IPTG addition. The CENP-T N terminus directly binds to the Ndc80 complex and the CENP-C N terminus binds to the Mis12 complex, which associates with the Ndc80 complex. Although both kinetochores do not contain CENP-A and CCAN, the CPC is recruited. If CENP-T or CENP-C N terminus were steadily supplied to the LacO locus, functional kinetochores would be formed. This indicates that CENP-T or CENP-C N terminus is sufficient for formation of functional kinetochores in the absence of CENP-A and most CCAN proteins. (B) Kinetochore formation at the LacO locus induced by LacI fusion with HJURP, CENP-I, full-length CENP-C, or CENP-C C terminus (601–864). These kinetochores are resistant to addition of IPTG, and full kinetochore components are recruited. HJURP is known as a CENP-A–specific chaperone. CENP-I and CENP-C C terminus function as a mark for CENP-A incorporation. Once a full kinetochore is formed at the LacO locus, Lac-I fusions with HJURP, CENP-I, or CENP-C proteins are dispensable. Characterization of this type of kinetochore suggests that a major function of CENP-C or CENP-I is recruitment of CENP-A.