TY - UNPB
T1 - Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation
AU - al-Rawi, Aymen
AU - Korolchuk, Svitlana
AU - Endicott, Jane
AU - Ly, Tony
N1 - The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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).
PY - 2022/5/24
Y1 - 2022/5/24
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 was 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 promote 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 was 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 promote 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 - Cell cycle
KW - Cyclin-dependent kinases
KW - Short linear motifs (SLiMs)
KW - Consensus sequence motifs
KW - Mass spectrometry
KW - Proteomics
KW - PTMs
U2 - 10.1101/2022.05.24.493195
DO - 10.1101/2022.05.24.493195
M3 - Preprint
BT - Cyclin A and Cks1 promote kinase consensus switching to non-proline directed CDK1 phosphorylation
PB - BioRxiv
CY - Cold Spring Harbour Laboratory
ER -