PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance

Pragya Parashara; Bethan Medina-Pritchard; Maria Alba Abad; Paula Sotelo-Parrilla; Reshma Thamkachy; David Grundei; Juan Zou; Christos Spanos; Chandni Natalia Kumar; Claire Basquin; Vimal Das; Zhaoyue Yan; Asma Abdullah Al-Murtadha; David A. Kelly; Toni Mchugh; Axel Imhof; Juri Rappsilber; Arockia A. Jeyaprakash

doi:10.1126/science.ado8270

PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance

Pragya Parashara, Bethan Medina-Pritchard, Maria Alba Abad, Paula Sotelo-Parrilla, Reshma Thamkachy, David Grundei, Juan Zou, Christos Spanos, Chandni Natalia Kumar, Claire Basquin, Vimal Das, Zhaoyue Yan, Asma Abdullah Al-Murtadha, David A. Kelly, Toni Mchugh, Axel Imhof, Juri Rappsilber, Arockia A. Jeyaprakash^*

^*Corresponding author for this work

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

Abstract / Description of output

Accurate chromosome segregation requires the attachment of microtubules to centromeres, epigenetically defined by the enrichment of CENP-A nucleosomes. During DNA replication, CENP-A nucleosomes undergo dilution. To preserve centromere identity, correct amounts of CENP-A must be restored in a cell-cycle controlled manner orchestrated by the Mis18 complex (Mis18α-Mis18β-Mis18BP1). We demonstrate that PLK1 interacts with Mis18 complex by recognising self-primed phosphorylations of Mis18α (Ser54) and Mis18BP1 (Thr78 and Ser93) through its Polo-Box domain. Disrupting these phosphorylations perturbed centromere recruitment of CENP-A chaperone HJURP and new CENP-A loading. Biochemical and functional analyses showed that phosphorylation of Mis18α and PLK1 binding were required to activate Mis18α-Mis18β and promote Mis18 complex-HJURP interaction. Thus, our study reveals key molecular events underpinning the licensing role of PLK1 in ensuring accurate centromere inheritance.

Original language	English
Pages (from-to)	1098–1104
Number of pages	7
Journal	Science
Volume	385
Issue number	6713
DOIs	https://doi.org/10.1126/science.ado8270
Publication status	Published - 6 Sept 2024

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The Hidden Cell Discovery Research Platform
Marston, A., Allshire, R., Davies, O., El Karoui, M., Kustatscher, G., O'Carroll, D., Rappsilber, J. & Rosser, S.
Wellcome Trust (Rcl)
1/12/23 → 30/11/30
Project: Research
Mechanistic Basis for CENP-32 Mediated Regulation of Cell Division
Arulanandam, J., Earnshaw, B. & Tollervey, D.
Medical Research Council
1/01/23 → 31/12/25
Project: Research
Core funding for the Wellcome Centre for Cell Biology
Marston, A.
Wellcome Trust (Rcl)
1/12/21 → 30/11/24
Project: Research

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Parashara, P., Medina-Pritchard, B., Alba Abad, M., Sotelo-Parrilla, P., Thamkachy, R., Grundei, D., Zou, J., Spanos, C., Kumar, C. N., Basquin, C., Das, V., Yan, Z., Al-Murtadha, A. A., Kelly, D. A., Mchugh, T., Imhof, A., Rappsilber, J., & Jeyaprakash, A. A. (2024). PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance. Science, 385(6713), 1098–1104. https://doi.org/10.1126/science.ado8270

@article{2edcaca422454af294e8c02def57a012,

title = "PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance",

abstract = "Accurate chromosome segregation requires the attachment of microtubules to centromeres, epigenetically defined by the enrichment of CENP-A nucleosomes. During DNA replication, CENP-A nucleosomes undergo dilution. To preserve centromere identity, correct amounts of CENP-A must be restored in a cell-cycle controlled manner orchestrated by the Mis18 complex (Mis18α-Mis18β-Mis18BP1). We demonstrate that PLK1 interacts with Mis18 complex by recognising self-primed phosphorylations of Mis18α (Ser54) and Mis18BP1 (Thr78 and Ser93) through its Polo-Box domain. Disrupting these phosphorylations perturbed centromere recruitment of CENP-A chaperone HJURP and new CENP-A loading. Biochemical and functional analyses showed that phosphorylation of Mis18α and PLK1 binding were required to activate Mis18α-Mis18β and promote Mis18 complex-HJURP interaction. Thus, our study reveals key molecular events underpinning the licensing role of PLK1 in ensuring accurate centromere inheritance. ",

author = "Pragya Parashara and Bethan Medina-Pritchard and {Alba Abad}, Maria and Paula Sotelo-Parrilla and Reshma Thamkachy and David Grundei and Juan Zou and Christos Spanos and Kumar, {Chandni Natalia} and Claire Basquin and Vimal Das and Zhaoyue Yan and Al-Murtadha, {Asma Abdullah} and Kelly, {David A.} and Toni Mchugh and Axel Imhof and Juri Rappsilber and Jeyaprakash, {Arockia A.}",

note = "We would like to thank the Wellcome Discovery Research Platform of Hidden Cell Biology, including the Centre for Optical Instrumentation Laboratory and the Proteomics Core for their help with microscopy, proteomics and analysis. We would also like to thank the Edinburgh Protein Production Facility. We would also like to acknowledge Diamond Light Source, where the crystal structure data was collected. In addition, we would like to thank Andrea Musacchio and Duccio Conti for discussion and sharing of unpublished data. ",

year = "2024",

month = sep,

day = "6",

doi = "10.1126/science.ado8270",

language = "English",

volume = "385",

pages = "1098–1104",

journal = "Science",

issn = "0036-8075",

publisher = "AMER ASSOC ADVANCEMENT SCIENCE",

number = "6713",

}

Parashara, P, Medina-Pritchard, B, Alba Abad, M, Sotelo-Parrilla, P, Thamkachy, R, Grundei, D, Zou, J, Spanos, C, Kumar, CN, Basquin, C, Das, V, Yan, Z, Al-Murtadha, AA, Kelly, DA , Mchugh, T, Imhof, A, Rappsilber, J & Jeyaprakash, AA 2024, 'PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance', Science, vol. 385, no. 6713, pp. 1098–1104. https://doi.org/10.1126/science.ado8270

TY - JOUR

T1 - PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance

AU - Parashara, Pragya

AU - Medina-Pritchard, Bethan

AU - Alba Abad, Maria

AU - Sotelo-Parrilla, Paula

AU - Thamkachy, Reshma

AU - Grundei, David

AU - Zou, Juan

AU - Spanos, Christos

AU - Kumar, Chandni Natalia

AU - Basquin, Claire

AU - Das, Vimal

AU - Yan, Zhaoyue

AU - Al-Murtadha, Asma Abdullah

AU - Kelly, David A.

AU - Mchugh, Toni

AU - Imhof, Axel

AU - Rappsilber, Juri

AU - Jeyaprakash, Arockia A.

N1 - We would like to thank the Wellcome Discovery Research Platform of Hidden Cell Biology, including the Centre for Optical Instrumentation Laboratory and the Proteomics Core for their help with microscopy, proteomics and analysis. We would also like to thank the Edinburgh Protein Production Facility. We would also like to acknowledge Diamond Light Source, where the crystal structure data was collected. In addition, we would like to thank Andrea Musacchio and Duccio Conti for discussion and sharing of unpublished data.

PY - 2024/9/6

Y1 - 2024/9/6

N2 - Accurate chromosome segregation requires the attachment of microtubules to centromeres, epigenetically defined by the enrichment of CENP-A nucleosomes. During DNA replication, CENP-A nucleosomes undergo dilution. To preserve centromere identity, correct amounts of CENP-A must be restored in a cell-cycle controlled manner orchestrated by the Mis18 complex (Mis18α-Mis18β-Mis18BP1). We demonstrate that PLK1 interacts with Mis18 complex by recognising self-primed phosphorylations of Mis18α (Ser54) and Mis18BP1 (Thr78 and Ser93) through its Polo-Box domain. Disrupting these phosphorylations perturbed centromere recruitment of CENP-A chaperone HJURP and new CENP-A loading. Biochemical and functional analyses showed that phosphorylation of Mis18α and PLK1 binding were required to activate Mis18α-Mis18β and promote Mis18 complex-HJURP interaction. Thus, our study reveals key molecular events underpinning the licensing role of PLK1 in ensuring accurate centromere inheritance.

AB - Accurate chromosome segregation requires the attachment of microtubules to centromeres, epigenetically defined by the enrichment of CENP-A nucleosomes. During DNA replication, CENP-A nucleosomes undergo dilution. To preserve centromere identity, correct amounts of CENP-A must be restored in a cell-cycle controlled manner orchestrated by the Mis18 complex (Mis18α-Mis18β-Mis18BP1). We demonstrate that PLK1 interacts with Mis18 complex by recognising self-primed phosphorylations of Mis18α (Ser54) and Mis18BP1 (Thr78 and Ser93) through its Polo-Box domain. Disrupting these phosphorylations perturbed centromere recruitment of CENP-A chaperone HJURP and new CENP-A loading. Biochemical and functional analyses showed that phosphorylation of Mis18α and PLK1 binding were required to activate Mis18α-Mis18β and promote Mis18 complex-HJURP interaction. Thus, our study reveals key molecular events underpinning the licensing role of PLK1 in ensuring accurate centromere inheritance.

U2 - 10.1126/science.ado8270

DO - 10.1126/science.ado8270

M3 - Article

SN - 0036-8075

VL - 385

SP - 1098

EP - 1104

JO - Science

JF - Science

IS - 6713

ER -

PLK1-Mediated Phosphorylation Cascade Activates Mis18 Complex to Ensure Centromere Inheritance

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The Hidden Cell Discovery Research Platform

Mechanistic Basis for CENP-32 Mediated Regulation of Cell Division

Core funding for the Wellcome Centre for Cell Biology

Cite this