A conserved CENP-E region mediates BubR1-independent recruitment to the outer corona at mitotic onset.

The outer corona plays an essential role at the onset of mitosis by expanding to maximize microtubule attachment to kinetochores. 1,2 The low-density structure of the corona forms through the expansion of unattached kinetochores. It comprises the RZZ complex, the dynein adaptor Spindly, the plus-end directed microtubule motor centromere protein E (CENP-E), and the Mad1/Mad2 spindle-assembly checkpoint proteins. 3,4,5,6,7,8,9,10 CENP-E specifically associates with unattached kinetochores to facilitate chromosome congression, 11,12,13,14,15,16 interacting with BubR1 at the kinetochore through its C-terminal region (2091–2358). 17,18,19,20,21 We recently showed that CENP-E recruitment to BubR1 at the kinetochores is both rapid and essential for correct chromosome alignment. However, CENP-E is also recruited to the outer corona by a second, slower pathway that is currently undefined. 19 Here, we show that BubR1-independent localization of CENP-E is mediated by a conserved loop that is essential for outer-corona targeting. We provide a structural model of the entire CENP-E kinetochore-targeting domain combining X-ray crystallography and Alphafold2. We reveal that maximal recruitment of CENP-E to unattached kinetochores critically depends on BubR1 and the outer corona, including dynein. Ectopic expression of the CENP-E C-terminal domain recruits the RZZ complex, Mad1, and Spindly, and prevents kinetochore biorientation in cells. We propose that BubR1-recruited CENP-E, in addition to its essential role in chromosome alignment to the metaphase plate, contributes to the recruitment of outer corona proteins through interactions with the CENP-E corona-targeting domain to facilitate the rapid capture of microtubules for efficient chromosome alignment and mitotic progression. [Display omitted] • CENP-E targets to unattached kinetochores through BubR1 and the RZZ complex/Spindly • CENP-E recruits the RZZ complex, Spindly, and Mad1 • Dynein is essential for both CENP-E targeting and removal Weber et al. demonstrate that CENP-E recruitment to kinetochores is dependent on both the BubR1- and the outer corona-dependent pathways and on dynein. They provide a molecular model of the CENP-E kinetochore-targeting domain and propose that CENP-E stabilizes the outer corona by recruiting the RZZ complex, Spindly and Mad1 to accelerate microtubule attachment. [ABSTRACT FROM AUTHOR]