The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface
During mitosis, the macromolecular kinetochore complex assembles on the centromere to orchestrate chromosome segregation. The properties and architecture of the 16-subunit Constitutive Centromere-Associated Network (CCAN) that allow it to build a robust platform for kinetochore assembly are poorly u...
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Elsevier
2017
|
| Online Access: | http://hdl.handle.net/1721.1/110167 https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 |
| _version_ | 1811080876256133120 |
|---|---|
| author | Sekulic, Nikolina Guo, Lucie Y. Black, Ben E. McKinley, Kara Lavidge Tsinman, Tonia Cheeseman, Iain M |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Sekulic, Nikolina Guo, Lucie Y. Black, Ben E. McKinley, Kara Lavidge Tsinman, Tonia Cheeseman, Iain M |
| author_sort | Sekulic, Nikolina |
| collection | MIT |
| description | During mitosis, the macromolecular kinetochore complex assembles on the centromere to orchestrate chromosome segregation. The properties and architecture of the 16-subunit Constitutive Centromere-Associated Network (CCAN) that allow it to build a robust platform for kinetochore assembly are poorly understood. Here, we use inducible CRISPR knockouts and biochemical reconstitutions to define the interactions between the human CCAN proteins. We find that the CCAN does not assemble as a linear hierarchy, and instead, each sub-complex requires multiple non-redundant interactions for its localization to centromeres and the structural integrity of the overall assembly. We demonstrate that the CENP-L-N complex plays a crucial role at the core of this assembly through interactions with CENP-C and CENP-H-I-K-M. Finally, we show that the CCAN is remodeled over the cell cycle such that sub-complexes depend on their interactions differentially. Thus, an interdependent meshwork within the CCAN underlies the centromere specificity and stability of the kinetochore. |
| first_indexed | 2024-09-23T11:38:17Z |
| format | Article |
| id | mit-1721.1/110167 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:38:17Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1101672022-09-27T20:57:29Z The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface Sekulic, Nikolina Guo, Lucie Y. Black, Ben E. McKinley, Kara Lavidge Tsinman, Tonia Cheeseman, Iain M Massachusetts Institute of Technology. Department of Biological Engineering Whitehead Institute for Biomedical Research McKinley, Kara Lavidge Tsinman, Tonia Cheeseman, Iain M During mitosis, the macromolecular kinetochore complex assembles on the centromere to orchestrate chromosome segregation. The properties and architecture of the 16-subunit Constitutive Centromere-Associated Network (CCAN) that allow it to build a robust platform for kinetochore assembly are poorly understood. Here, we use inducible CRISPR knockouts and biochemical reconstitutions to define the interactions between the human CCAN proteins. We find that the CCAN does not assemble as a linear hierarchy, and instead, each sub-complex requires multiple non-redundant interactions for its localization to centromeres and the structural integrity of the overall assembly. We demonstrate that the CENP-L-N complex plays a crucial role at the core of this assembly through interactions with CENP-C and CENP-H-I-K-M. Finally, we show that the CCAN is remodeled over the cell cycle such that sub-complexes depend on their interactions differentially. Thus, an interdependent meshwork within the CCAN underlies the centromere specificity and stability of the kinetochore. United States. National Institutes of Health (GM088313) United States. National Institutes of Health (GM108718) American Cancer Society (121776) 2017-06-22T15:14:56Z 2017-06-22T15:14:56Z 2015-11 2015-09 Article http://purl.org/eprint/type/JournalArticle 1097-2765 1097-4164 http://hdl.handle.net/1721.1/110167 McKinley, Kara L.; Sekulic, Nikolina; Guo, Lucie Y.; Tsinman, Tonia; Black, Ben E. and Cheeseman, Iain M. “The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface.” Molecular Cell 60, no. 6 (December 2015): 886–898 © 2015 Elsevier Inc https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 en_US http://dx.doi.org/10.1016/j.molcel.2015.10.027 Molecular Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Sekulic, Nikolina Guo, Lucie Y. Black, Ben E. McKinley, Kara Lavidge Tsinman, Tonia Cheeseman, Iain M The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title | The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title_full | The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title_fullStr | The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title_full_unstemmed | The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title_short | The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface |
| title_sort | cenp l n complex forms a critical node in an integrated meshwork of interactions at the centromere kinetochore interface |
| url | http://hdl.handle.net/1721.1/110167 https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 |
| work_keys_str_mv | AT sekulicnikolina thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT guoluciey thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT blackbene thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT mckinleykaralavidge thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT tsinmantonia thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT cheesemaniainm thecenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT sekulicnikolina cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT guoluciey cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT blackbene cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT mckinleykaralavidge cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT tsinmantonia cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface AT cheesemaniainm cenplncomplexformsacriticalnodeinanintegratedmeshworkofinteractionsatthecentromerekinetochoreinterface |