Human Ska complex and Ndc80 complex interact to form a load-bearing assembly that strengthens kinetochore-microtubule attachments

被引:62
作者
Helgeson, Luke A. [1 ]
Zelter, Alex [1 ]
Riffle, Michael [1 ]
MacCoss, Michael J. [2 ]
Asbury, Charles L. [3 ]
Davis, Trisha N. [1 ]
机构
[1] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
[2] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA
[3] Univ Washington, Dept Physiol & Biophys, Seattle, WA 98195 USA
关键词
kinetochore; microtubules; optical tweezers; Ska complex; Ndc80; complex; MITOTIC PROGRESSION; OUTER PLATE; AURORA B; BINDING; DAM1; COMPONENT; DYNAMICS; NETWORK; ORGANIZATION; INTERFACE;
D O I
10.1073/pnas.1718553115
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Accurate segregation of chromosomes relies on the force-bearing capabilities of the kinetochore to robustly attach chromosomes to dynamic microtubule tips. The human Ska complex and Ndc80 complex are outer-kinetochore components that bind microtubules and are required to fully stabilize kinetochore-microtubule attachments in vivo. While purified Ska complex tracks with disassembling microtubule tips, it remains unclear whether the Ska complex-microtubule interaction is sufficiently strong to make a significant contribution to kinetochore-microtubule coupling. Alternatively, Ska complex might affect kinetochore coupling indirectly, through recruitment of phosphoregulatory factors. Using optical tweezers, we show that the Ska complex itself bears load on microtubule tips, strengthens Ndc80 complex-based tip attachments, and increases the switching dynamics of the attached microtubule tips. Cross-linking mass spectrometry suggests the Ska complex directly binds Ndc80 complex through interactions between the Ska3 unstructured C-terminal region and the coiled-coil regions of each Ndc80 complex subunit. Deletion of the Ska complex microtubule-binding domain or the Ska3 C terminus prevents Ska complex from strengthening Ndc80 complex-based attachments. Together, our results indicate that the Ska complex can directly strengthen the kinetochore-microtubule interface and regulate microtubule tip dynamics by forming an additional connection between the Ndc80 complex and the microtubule.
引用
收藏
页码:2740 / 2745
页数:6
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