Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation

Aymen Al-Rawi; Edward Kaye; Svitlana Korolchuk; Jane A. Endicott; Tony Ly

doi:10.1016/j.celrep.2023.112139

Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation

Aymen Al-Rawi, Edward Kaye, Svitlana Korolchuk, Jane A. Endicott, Tony Ly (Lead / Corresponding author)

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

4 Citations (Scopus)

325 Downloads (Pure)

Abstract

Ordered protein phosphorylation by CDKs is a key mechanism for regulating the cell cycle. How temporal order is enforced in mammalian cells remains unclear. Using a fixed cell kinase assay and phosphoproteomics, we show how CDK1 activity and non-catalytic CDK1 subunits contribute to the choice of substrate and site of phosphorylation. Increases in CDK1 activity alters substrate choice, with intermediate and low sensitivity CDK1 substrates enriched in DNA replication and mitotic functions, respectively. This activity dependence is shared between Cyclin A- and Cyclin B-CDK1. Cks1 has a proteome-wide role as an enhancer of multisite CDK1 phosphorylation. Contrary to the model of CDK1 as an exclusively proline-directed kinase, we show that Cyclin A and Cks1 enhance non-proline directed phosphorylation, preferably on sites with a +3 lysine residue. Indeed, 70% of cell cycle regulated phosphorylations, where the kinase carrying out this modification has not been identified, are non-proline directed CDK1 sites.

Original language	English
Article number	112139
Number of pages	21
Journal	Cell Reports
Volume	42
Issue number	3
Early online date	24 Feb 2023
DOIs	https://doi.org/10.1016/j.celrep.2023.112139
Publication status	Published - 28 Mar 2023

Keywords

CP: Cell biology
CP: Molecular biology
cell cycle
kinase consensus motif
mitosis
phosphoproteomics

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology

Access to Document

10.1016/j.celrep.2023.112139Licence: CC BY

Final Published VersionFinal published version, 9.57 MBLicence: CC BY

Cite this

@article{c7e54fde1d07492595e2ec9887c1b656,

title = "Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation",

abstract = "Ordered protein phosphorylation by CDKs is a key mechanism for regulating the cell cycle. How temporal order is enforced in mammalian cells remains unclear. Using a fixed cell kinase assay and phosphoproteomics, we show how CDK1 activity and non-catalytic CDK1 subunits contribute to the choice of substrate and site of phosphorylation. Increases in CDK1 activity alters substrate choice, with intermediate and low sensitivity CDK1 substrates enriched in DNA replication and mitotic functions, respectively. This activity dependence is shared between Cyclin A- and Cyclin B-CDK1. Cks1 has a proteome-wide role as an enhancer of multisite CDK1 phosphorylation. Contrary to the model of CDK1 as an exclusively proline-directed kinase, we show that Cyclin A and Cks1 enhance non-proline directed phosphorylation, preferably on sites with a +3 lysine residue. Indeed, 70% of cell cycle regulated phosphorylations, where the kinase carrying out this modification has not been identified, are non-proline directed CDK1 sites.",

keywords = "CP: Cell biology, CP: Molecular biology, cell cycle, kinase consensus motif, mitosis, phosphoproteomics",

author = "Aymen Al-Rawi and Edward Kaye and Svitlana Korolchuk and Endicott, {Jane A.} and Tony Ly",

note = "Funding Information: This work is supported by a Wellcome Trust and Royal Society Sir Henry Dale Fellowship to T.L. (206211/Z/17/Z), a Darwin Trust PhD studentship to A. A., a UK Medical Research Council Programme Grant to J.E. (MR/N009738/1), core funding for the Wellcome Centre for Cell Biology (091020), a Wellcome Multi-User Equipment Grant to T.L. (218305/Z/19/Z) and a Wellcome Innovation Award for mass spectrometry equipment (218448/Z/19/Z). Copyright: {\textcopyright} 2023, The Authors.",

year = "2023",

month = mar,

day = "28",

doi = "10.1016/j.celrep.2023.112139",

language = "English",

volume = "42",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Elsevier",

number = "3",

}

TY - JOUR

T1 - Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation

AU - Al-Rawi, Aymen

AU - Kaye, Edward

AU - Korolchuk, Svitlana

AU - Endicott, Jane A.

AU - Ly, Tony

N1 - Funding Information: This work is supported by a Wellcome Trust and Royal Society Sir Henry Dale Fellowship to T.L. (206211/Z/17/Z), a Darwin Trust PhD studentship to A. A., a UK Medical Research Council Programme Grant to J.E. (MR/N009738/1), core funding for the Wellcome Centre for Cell Biology (091020), a Wellcome Multi-User Equipment Grant to T.L. (218305/Z/19/Z) and a Wellcome Innovation Award for mass spectrometry equipment (218448/Z/19/Z). Copyright: © 2023, The Authors.

PY - 2023/3/28

Y1 - 2023/3/28

N2 - Ordered protein phosphorylation by CDKs is a key mechanism for regulating the cell cycle. How temporal order is enforced in mammalian cells remains unclear. Using a fixed cell kinase assay and phosphoproteomics, we show how CDK1 activity and non-catalytic CDK1 subunits contribute to the choice of substrate and site of phosphorylation. Increases in CDK1 activity alters substrate choice, with intermediate and low sensitivity CDK1 substrates enriched in DNA replication and mitotic functions, respectively. This activity dependence is shared between Cyclin A- and Cyclin B-CDK1. Cks1 has a proteome-wide role as an enhancer of multisite CDK1 phosphorylation. Contrary to the model of CDK1 as an exclusively proline-directed kinase, we show that Cyclin A and Cks1 enhance non-proline directed phosphorylation, preferably on sites with a +3 lysine residue. Indeed, 70% of cell cycle regulated phosphorylations, where the kinase carrying out this modification has not been identified, are non-proline directed CDK1 sites.

AB - Ordered protein phosphorylation by CDKs is a key mechanism for regulating the cell cycle. How temporal order is enforced in mammalian cells remains unclear. Using a fixed cell kinase assay and phosphoproteomics, we show how CDK1 activity and non-catalytic CDK1 subunits contribute to the choice of substrate and site of phosphorylation. Increases in CDK1 activity alters substrate choice, with intermediate and low sensitivity CDK1 substrates enriched in DNA replication and mitotic functions, respectively. This activity dependence is shared between Cyclin A- and Cyclin B-CDK1. Cks1 has a proteome-wide role as an enhancer of multisite CDK1 phosphorylation. Contrary to the model of CDK1 as an exclusively proline-directed kinase, we show that Cyclin A and Cks1 enhance non-proline directed phosphorylation, preferably on sites with a +3 lysine residue. Indeed, 70% of cell cycle regulated phosphorylations, where the kinase carrying out this modification has not been identified, are non-proline directed CDK1 sites.

KW - CP: Cell biology

KW - CP: Molecular biology

KW - cell cycle

KW - kinase consensus motif

KW - mitosis

KW - phosphoproteomics

UR - http://www.scopus.com/inward/record.url?scp=85148671580&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2023.112139

DO - 10.1016/j.celrep.2023.112139

M3 - Article

C2 - 36840943

SN - 2211-1247

VL - 42

JO - Cell Reports

JF - Cell Reports

IS - 3

M1 - 112139

ER -

Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

A Platform for Drug Target Deconvolution and Exploitation

Understanding the Proliferation-Quiescence Switch Using Quantitative Cellular Biochemistry (Sir Henry Dale Fellowship) (Transfer from University of Edinburgh)