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Open AccessOpen Access JUSTC Life Sciences 15 May 2023

The function, structure and dynamic organization of centromeres and kinetochores

  • 1.

    MOE Key Laboratory for Cellular Dynamics, School of Life Sciences, University of Science and Technology of China, Hefei 230026, China

  • 2.

    Hefei National Research Center for Physical Sciences at the Microscale, Anhui Key Laboratory for Cellular Dynamics and Chemical Biology, University of Science and Technology of China, Hefei 230027, China

Cite this:
https://doi.org/10.52396/JUSTC-2022-0184
More Information
  • Author Bio:

    Zhen Dou received his Ph.D. degree in Cell Biology from the University of Science and Technology of China. He is currently an Associate Professor at the University of Science and Technology of China. His research interests include cell division cycle and genome stability maintenance

    Xing Liu is a Professor of MOE Key Laboratory for Cellular Dynamics at the University of Science and Technology of China. She received her Ph.D. degree in Cell Biology from the University of Science and Technology of China. Her research mainly focuses on solving the key scientific questions and challenges of cell fate decision using chemical tools and methods

  • Corresponding author: E-mail: douzhen@ustc.edu.cn; E-mail: xing1017@ustc.edu.cn
  • Received Date: 27 December 2022
  • Accepted Date: 07 April 2023
    • Available Online: 15 May 2023
    Abstract

  • Abstract

    It is a fundamental task to ensure the faithful transmission of genetic information across generations for eukaryote species. The centromere is a specialized chromosomal region that is essential for mediating sister chromatid alignment and separation during mitosis. Centromere identity is epigenetically determined by nucleosome-containing centromere protein A (CENP-A). The CENP-A nucleosome provides the foundation for the association of the inner kinetochore and the assembly of the outer kinetochore in mitosis. Here we review centromere identity determination, inner kinetochore function and assembly, and outer kinetochore function and assembly. In particular, we focus on the recent advances in the structure-activity relationship of the constitutive centromere-associated network (CCAN). CCAN structure information sheds new light on our understanding of centromere and kinetochore functions and dynamic organization.

    Graphical abstract

    Schematic representation of a mitotic spindle and the organization of core proteins of inner and outer kinetochore.

    Abstract

    It is a fundamental task to ensure the faithful transmission of genetic information across generations for eukaryote species. The centromere is a specialized chromosomal region that is essential for mediating sister chromatid alignment and separation during mitosis. Centromere identity is epigenetically determined by nucleosome-containing centromere protein A (CENP-A). The CENP-A nucleosome provides the foundation for the association of the inner kinetochore and the assembly of the outer kinetochore in mitosis. Here we review centromere identity determination, inner kinetochore function and assembly, and outer kinetochore function and assembly. In particular, we focus on the recent advances in the structure-activity relationship of the constitutive centromere-associated network (CCAN). CCAN structure information sheds new light on our understanding of centromere and kinetochore functions and dynamic organization.

    • Public Summary

    • Centromere is a vital macromolecular machinery for cell renewal quality control.
    • CENP-A nucleosome is the hallmark and foundation for centromere and kinetochore assembly.
    • Centromere and kinetochore core complex CCAN undergoes dynamic remodeling from CENP-A nucleosome-elicited loading to DNA-centric gripping upon assembly.
    • The excitement and challenge ahead are to illuminate the spatiotemporal characteristics of CCAN assembly in situ at atomic resolution.

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    • References(79)
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    Figure  1.  A cartoon shown mitotic spindle and the chromosomes. In the central position of the spindle, two chromosomes achieved correct microtubule attachment. On the contrary, a chromosome near the left pole failed to establish proper microtubule attachment. Thus, the kinetochores of this unaligned chromosome initiate a signaling call the spindle assembly checkpoint (SAC) and thereafter preclude the cell to enter anaphase. The yellow dots on chromosomes depict centromeres, and the red dots depict kinetochores.

    Figure  2.  The structure of inner kinetochore and outer kinetochore core proteins. Centromeric CENP-A nucleosome serves as the foundation for kinetochore assembly. CCAN, a complex comprising 16 proteins, constitutively localizes at the centromeres. The cartoon shows the 3D architecture of CCAN bound CENP-A nucleosome. Interestingly, CCAN forms a channel that topologically grasps linker DNA of CENP-A nucleosome. CENP-T and CENP-C, which both have an elongated N-terminal tail, functions as two parallel pathways to recruit Knl1 complex, Mis12 complex and Ndc80 complex in mitosis. Knl1 complex, Mis12 complex and Ndc80 complex comprise the core of outer kinetochore and were named KMN network. KMN harbors two activity, attaching the spindle microtubule and recruiting SAC signaling proteins.

    Figure  3.  DNA binds to CCAN through the CENP-LN channel[32]. (a) Electrostatic potential surface view of CENP-LN-HIK head-TW binding with DNA. The DNA is shown as cartoon. Note that positively charged amino acids from CENPLN, CENP-I and CENP-TW constitute the contact sites between CCAN and DNA. (b) Representative immunofluorescence montage of HeLa cells expressing GFPCENP-L wild type and DNA binding-deficient mutants. 4KA represents CENP-L K155A/R306A/K319A/K321A, 4KE represents K155E/R306E/K319E/K321E. Scale bar, 10 μm. (c) Statistical analysis of kinetochore intensity of various GFP-CENP-L mutants as treated in b. Bars represent the mean kinetochore intensity (±SEM) normalized to values of the CENP-L WT expressing group. Each dot represents one cell (30 cells from three independent experiments). Ordinary one-way ANOVA followed by Tukey’s post hoc test was used to determine statistical significance. ****p < 0.0001.

    [1]
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    [2]
    Biggins S. The composition, functions, and regulation of the budding yeast kinetochore. Genetics, 2013, 194 (4): 817–846. doi: 10.1534/genetics.112.145276
    [3]
    Lawrimore J, Bloom K S, Salmon E D. Point centromeres contain more than a single centromere-specific Cse4 (CENP-A) nucleosome. J. Cell Biol., 2011, 195 (4): 573–582. doi: 10.1083/jcb.201106036
    [4]
    Coffman V C, Wu P C, Parthun M R, et al. CENP-A exceeds microtubule attachment sites in centromere clusters of both budding and fission yeast. J. Cell Biol., 2011, 195 (4): 563–572. doi: 10.1083/jcb.201106078
    [5]
    Haase J, Mishra P K, Stephens A, et al. A 3D map of the yeast kinetochore reveals the presence of core and accessory centromere-specific histone. Curr. Biol., 2013, 23 (19): 1939–1944. doi: 10.1016/j.cub.2013.07.083
    [6]
    Cieslinski K, Wu Y L, Nechyporenko L, et al. Nanoscale structural organization and stoichiometry of the budding yeast kinetochore. J. Cell Biol., 2023, 222 (4): e202209094. doi: 10.1083/jcb.202209094
    [7]
    Cleveland D W, Mao Y, Sullivan K F. Centromeres and kinetochores: From epigenetics to mitotic checkpoint signaling. Cell, 2003, 112 (4): 407–421. doi: 10.1016/S0092-8674(03)00115-6
    [8]
    Allshire R C, Karpen G H. Epigenetic regulation of centromeric chromatin: old dogs, new tricks? Nat. Rev. Genet., 2008, 9 (12): 923–937. doi: 10.1038/nrg2466
    [9]
    Steiner F A, Henikoff S. Holocentromeres are dispersed point centromeres localized at transcription factor hotspots. eLife, 2014, 3: e02025. doi: 10.7554/eLife.02025
    [10]
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