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

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  • Jeraldine Weber
  • Thibault Legal
  • Alicia Perez Lezcano
  • Agata Gluszek-Kustusz
  • Calum Paterson
  • Susana Eibes
  • Barisic, Marin
  • Owen R. Davies
  • Julie P.I. Welburn
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.
OriginalsprogEngelsk
TidsskriftCurrent Biology
Vol/bind34
Udgave nummer5
Sider (fra-til)1133-1141.e4
Antal sider9
ISSN0960-9822
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
We thank I. Chambers and R. Pantier for pENTR-GFP and E. Campeau for pLenti CMV/TO-DEST. We thank Jingchao Wu, Geert Kops, Verena Cmentowski, and Andrea Musacchio for sharing data and information before publication. We thank Toni McHugh and Dave Kelly for support in the COIL imaging facility and HY Kok for help with cloning. We thank Life Science Editors for their help with manuscript editing. M. Barisic is supported by the Novo Nordisk Foundation ( NNF19OC0058504 ) and the Danish Council for Independent Research ( DFF-3101-00075 ). J.W. and O.D. are supported by a Wellcome Senior Research Fellowship ( 207430 and 219413 respectively). J.W. is also a EMBO Young Investigator. The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust ( 203149 ).

Funding Information:
We thank I. Chambers and R. Pantier for pENTR-GFP and E. Campeau for pLenti CMV/TO-DEST. We thank Jingchao Wu, Geert Kops, Verena Cmentowski, and Andrea Musacchio for sharing data and information before publication. We thank Toni McHugh and Dave Kelly for support in the COIL imaging facility and HY Kok for help with cloning. We thank Life Science Editors for their help with manuscript editing. M. Barisic is supported by the Novo Nordisk Foundation (NNF19OC0058504) and the Danish Council for Independent Research (DFF-3101-00075). J.W. and O.D. are supported by a Wellcome Senior Research Fellowship (207430 and 219413 respectively). J.W. is also a EMBO Young Investigator. The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust (203149). Conceptualization, J.W. T.L. And J.W.C. Investigation and formal analysis, J.W.C. T.L. A.L.P. A.G. C.P. O.D. S.E. and J.W. Writing – original fraft, J.W. Writing – review & editing, J.W. O.D. T.L. J.W.C. A.L.P. A.G. and M.B. Funding acquisition, J.W. O.D. and M.B. Resources, J.W. and M.B. Supervision, J.W. The authors declare no competing interests.

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© 2024 The Author(s)

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