Structural basis for Mis18 complex assembly and its implications for centromere maintenance

Reshma Thamkachy; Bethan Medina-Pritchard; Sang Ho Park; Carla G Chiodi; Juan Zou; Maria de la Torre-Barranco; Kazuma Shimanaka; Maria Alba Abad; Cristina Gallego Páramo; Regina Feederle; Emilija Ruksenaite; Patrick Heun; Owen R Davies; Juri Rappsilber; Dina Schneidman-Duhovny; Uhn-Soo Cho; A. Arockia Jeyaprakash

doi:10.1038/s44319-024-00183-w

Structural basis for Mis18 complex assembly and its implications for centromere maintenance

Reshma Thamkachy, Bethan Medina-Pritchard, Sang Ho Park, Carla G Chiodi, Juan Zou, Maria de la Torre-Barranco, Kazuma Shimanaka, Maria Alba Abad, Cristina Gallego Páramo, Regina Feederle, Emilija Ruksenaite, Patrick Heun, Owen R Davies, Juri Rappsilber, Dina Schneidman-Duhovny, Uhn-Soo Cho, A. Arockia Jeyaprakash^*

^*Corresponding author for this work

School of Biological Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

The centromere, defined by the enrichment of CENP-A (a Histone H3 variant) containing nucleosomes, is a specialised chromosomal locus that acts as a microtubule attachment site. To preserve centromere identity, CENP-A levels must be maintained through active CENP-A loading during the cell cycle. A central player mediating this process is the Mis18 complex (Mis18α, Mis18β and Mis18BP1), which recruits the CENP-A-specific chaperone HJURP to centromeres for CENP-A deposition. Here, using a multi-pronged approach, we characterise the structure of the Mis18 complex and show that multiple hetero- and homo-oligomeric interfaces facilitate the hetero-octameric Mis18 complex assembly composed of 4 Mis18α, 2 Mis18β and 2 Mis18BP1. Evaluation of structure-guided/separation-of-function mutants reveals structural determinants essential for cell cycle controlled Mis18 complex assembly and centromere maintenance. Our results provide new mechanistic insights on centromere maintenance, highlighting that while Mis18α can associate with centromeres and deposit CENP-A independently of Mis18β, the latter is indispensable for the optimal level of CENP-A loading required for preserving the centromere identity.

Original language	English
Pages (from-to)	3348-3372
Number of pages	25
Journal	EMBO Reports
Volume	25
Issue number	8
Early online date	1 Jul 2024
DOIs	https://doi.org/10.1038/s44319-024-00183-w
Publication status	Published - 9 Aug 2024

Keywords / Materials (for Non-textual outputs)

Centromere
CENP-A
Mis18 Complex
Centromere Inheritance
Integrative Structural Analysis

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10.1038/s44319-024-00183-w

thamkachy-et-al-2024-structural-basis-for-mis18-complex-assembly-and-its-implications-for-centromere-maintenanceFinal published version, 6.92 MBLicence: Creative Commons: Attribution (CC-BY)

1 Active
6 Finished

Molecular basis of chromosome synapsis and genetic exchange in mammalian meiosis
Davies, O. (Principal Investigator)
UK-based charities
15/02/21 → 14/02/26
Project: Research
Core funding for the Wellcome Centre for Cell Biology
Marston, A. (Principal Investigator)
Wellcome Trust (Rcl)
1/12/21 → 30/11/24
Project: Research
Molecular Mechanisms of Centromere Inheritance and Kinetochore Function
Arulanandam, J. (Principal Investigator)
Wellcome Trust (Rcl)
4/01/17 → 31/12/22
Project: Research

Cite this

Thamkachy, R., Medina-Pritchard, B., Park, S. H., Chiodi, C. G., Zou, J., de la Torre-Barranco, M., Shimanaka, K., Abad, M. A., Gallego Páramo, C., Feederle, R., Ruksenaite, E., Heun, P., Davies, O. R., Rappsilber, J., Schneidman-Duhovny, D., Cho, U.-S., & Jeyaprakash, A. A. (2024). Structural basis for Mis18 complex assembly and its implications for centromere maintenance. EMBO Reports, 25(8), 3348-3372. https://doi.org/10.1038/s44319-024-00183-w

@article{c54c52b3070948e1924b0244c99c320f,

title = "Structural basis for Mis18 complex assembly and its implications for centromere maintenance",

abstract = "The centromere, defined by the enrichment of CENP-A (a Histone H3 variant) containing nucleosomes, is a specialised chromosomal locus that acts as a microtubule attachment site. To preserve centromere identity, CENP-A levels must be maintained through active CENP-A loading during the cell cycle. A central player mediating this process is the Mis18 complex (Mis18α, Mis18β and Mis18BP1), which recruits the CENP-A-specific chaperone HJURP to centromeres for CENP-A deposition. Here, using a multi-pronged approach, we characterise the structure of the Mis18 complex and show that multiple hetero- and homo-oligomeric interfaces facilitate the hetero-octameric Mis18 complex assembly composed of 4 Mis18α, 2 Mis18β and 2 Mis18BP1. Evaluation of structure-guided/separation-of-function mutants reveals structural determinants essential for cell cycle controlled Mis18 complex assembly and centromere maintenance. Our results provide new mechanistic insights on centromere maintenance, highlighting that while Mis18α can associate with centromeres and deposit CENP-A independently of Mis18β, the latter is indispensable for the optimal level of CENP-A loading required for preserving the centromere identity.",

keywords = "Centromere, CENP-A, Mis18 Complex, Centromere Inheritance, Integrative Structural Analysis",

author = "Reshma Thamkachy and Bethan Medina-Pritchard and Park, {Sang Ho} and Chiodi, {Carla G} and Juan Zou and {de la Torre-Barranco}, Maria and Kazuma Shimanaka and Abad, {Maria Alba} and {Gallego P{\'a}ramo}, Cristina and Regina Feederle and Emilija Ruksenaite and Patrick Heun and Davies, {Owen R} and Juri Rappsilber and Dina Schneidman-Duhovny and Uhn-Soo Cho and Jeyaprakash, {A. Arockia}",

note = "The authors would like to thank David Kelly from the Centre Optical Instrumentation Laboratory, Martin Wear from the Edinburgh Protein Production Facility as well as Marcus Wilson and Maarten Tuijtel of the Cryo-Electron Microscopy Facility for their help. The authors also thank Diamond Light Source and the staff of beamline B21 (proposal sm23510), as well as Advanced Photon Source and the staff at beamlines LS-CAT 21 ID-G and ID-D. Thanks also to Iain Cheeseman for the kind gift of cell lines. ",

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month = aug,

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doi = "10.1038/s44319-024-00183-w",

language = "English",

volume = "25",

pages = "3348--3372",

journal = "EMBO Reports",

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Thamkachy, R, Medina-Pritchard, B, Park, SH, Chiodi, CG, Zou, J, de la Torre-Barranco, M, Shimanaka, K, Abad, MA, Gallego Páramo, C, Feederle, R, Ruksenaite, E, Heun, P, Davies, OR , Rappsilber, J, Schneidman-Duhovny, D, Cho, U-S & Jeyaprakash, AA 2024, 'Structural basis for Mis18 complex assembly and its implications for centromere maintenance', EMBO Reports, vol. 25, no. 8, pp. 3348-3372. https://doi.org/10.1038/s44319-024-00183-w

TY - JOUR

T1 - Structural basis for Mis18 complex assembly and its implications for centromere maintenance

AU - Thamkachy, Reshma

AU - Medina-Pritchard, Bethan

AU - Park, Sang Ho

AU - Chiodi, Carla G

AU - Zou, Juan

AU - de la Torre-Barranco, Maria

AU - Shimanaka, Kazuma

AU - Abad, Maria Alba

AU - Gallego Páramo, Cristina

AU - Feederle, Regina

AU - Ruksenaite, Emilija

AU - Heun, Patrick

AU - Davies, Owen R

AU - Rappsilber, Juri

AU - Schneidman-Duhovny, Dina

AU - Cho, Uhn-Soo

AU - Jeyaprakash, A. Arockia

N1 - The authors would like to thank David Kelly from the Centre Optical Instrumentation Laboratory, Martin Wear from the Edinburgh Protein Production Facility as well as Marcus Wilson and Maarten Tuijtel of the Cryo-Electron Microscopy Facility for their help. The authors also thank Diamond Light Source and the staff of beamline B21 (proposal sm23510), as well as Advanced Photon Source and the staff at beamlines LS-CAT 21 ID-G and ID-D. Thanks also to Iain Cheeseman for the kind gift of cell lines.

PY - 2024/8/9

Y1 - 2024/8/9

N2 - The centromere, defined by the enrichment of CENP-A (a Histone H3 variant) containing nucleosomes, is a specialised chromosomal locus that acts as a microtubule attachment site. To preserve centromere identity, CENP-A levels must be maintained through active CENP-A loading during the cell cycle. A central player mediating this process is the Mis18 complex (Mis18α, Mis18β and Mis18BP1), which recruits the CENP-A-specific chaperone HJURP to centromeres for CENP-A deposition. Here, using a multi-pronged approach, we characterise the structure of the Mis18 complex and show that multiple hetero- and homo-oligomeric interfaces facilitate the hetero-octameric Mis18 complex assembly composed of 4 Mis18α, 2 Mis18β and 2 Mis18BP1. Evaluation of structure-guided/separation-of-function mutants reveals structural determinants essential for cell cycle controlled Mis18 complex assembly and centromere maintenance. Our results provide new mechanistic insights on centromere maintenance, highlighting that while Mis18α can associate with centromeres and deposit CENP-A independently of Mis18β, the latter is indispensable for the optimal level of CENP-A loading required for preserving the centromere identity.

AB - The centromere, defined by the enrichment of CENP-A (a Histone H3 variant) containing nucleosomes, is a specialised chromosomal locus that acts as a microtubule attachment site. To preserve centromere identity, CENP-A levels must be maintained through active CENP-A loading during the cell cycle. A central player mediating this process is the Mis18 complex (Mis18α, Mis18β and Mis18BP1), which recruits the CENP-A-specific chaperone HJURP to centromeres for CENP-A deposition. Here, using a multi-pronged approach, we characterise the structure of the Mis18 complex and show that multiple hetero- and homo-oligomeric interfaces facilitate the hetero-octameric Mis18 complex assembly composed of 4 Mis18α, 2 Mis18β and 2 Mis18BP1. Evaluation of structure-guided/separation-of-function mutants reveals structural determinants essential for cell cycle controlled Mis18 complex assembly and centromere maintenance. Our results provide new mechanistic insights on centromere maintenance, highlighting that while Mis18α can associate with centromeres and deposit CENP-A independently of Mis18β, the latter is indispensable for the optimal level of CENP-A loading required for preserving the centromere identity.

KW - Centromere

KW - CENP-A

KW - Mis18 Complex

KW - Centromere Inheritance

KW - Integrative Structural Analysis

U2 - 10.1038/s44319-024-00183-w

DO - 10.1038/s44319-024-00183-w

M3 - Article

C2 - 38951710

SN - 1469-221X

VL - 25

SP - 3348

EP - 3372

JO - EMBO Reports

JF - EMBO Reports

IS - 8

ER -

Structural basis for Mis18 complex assembly and its implications for centromere maintenance

Abstract

Keywords / Materials (for Non-textual outputs)

Access to Document

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Projects

Molecular basis of chromosome synapsis and genetic exchange in mammalian meiosis

Core funding for the Wellcome Centre for Cell Biology

Molecular Mechanisms of Centromere Inheritance and Kinetochore Function

Cite this