Dataset Information

Cell cycle phosphorylation dynamics in vivo reveals switch-like control of intrinsic disorder

ABSTRACT: Switch-like cyclin-dependent kinase (CDK)-1 activation is thought to underlie the abruptness of mitotic onset, but how CDKs can simultaneously phosphorylate many diverse substrates is unknown, and direct evidence for such phosphorylation dynamics in vivo is lacking. Here, we analysed protein phosphorylation states in single Xenopus embryos throughout synchronous cell cycles. 22% of 4583 phosphosites on 1843 proteins were dynamic in vivo. We assigned cell cycle phases using egg extracts, showing 693 S-phase and 1035 mitotic phosphorylations. High-time resolution targeted phosphoproteomics in single embryos revealed switch-like mitotic phosphorylation of diverse protein complexes. 60% of cell cycle-regulated phosphosites occurred in CDK consensus motifs, and 72% located to intrinsically disordered regions (IDRs). Dynamically phosphorylated proteins, and documented CDK substrates in human and yeast, are significantly more disordered than targets of other cell cycle kinases and phosphoproteins in general. Furthermore, 30-50% are components of membraneless organelles. Our results suggest that CDK-mediated phosphorylation of intrinsic disorder allows switch-like mitotic cellular reorganisation.

ORGANISM(S): Xenopus Laevis

SUBMITTER: Juan Manuel Valverde

PROVIDER: PXD026088 | panorama | Fri Oct 13 00:00:00 BST 2023

REPOSITORIES: PanoramaPublic

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Publications

A cyclin-dependent kinase-mediated phosphorylation switch of disordered protein condensation.

Valverde Juan Manuel JM Dubra Geronimo G Phillips Michael M Haider Austin A Elena-Real Carlos C Fournet Aurélie A Alghoul Emile E Chahar Dhanvantri D Andrés-Sanchez Nuria N Paloni Matteo M Bernadó Pau P van Mierlo Guido G Vermeulen Michiel M van den Toorn Henk H Heck Albert J R AJR Constantinou Angelos A Barducci Alessandro A Ghosh Kingshuk K Sibille Nathalie N Knipscheer Puck P Krasinska Liliana L Fisher Daniel D Altelaar Maarten M

Nature communications 20231009 1

Cell cycle transitions result from global changes in protein phosphorylation states triggered by cyclin-dependent kinases (CDKs). To understand how this complexity produces an ordered and rapid cellular reorganisation, we generated a high-resolution map of changing phosphosites throughout unperturbed early cell cycles in single Xenopus embryos, derived the emergent principles through systems biology analysis, and tested them by biophysical modelling and biochemical experiments. We found that mos ...[more]

PMID: 37813838

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Project description:Minichromosome maintenance protein 2 (MCM2) is a licensing factor for DNA replication. It interacts with other MCM proteins to comprise MCM2-7 complex, which acts as a helicase for DNA unwinding and limits DNA replication to one round per cell cycle. MCM2 has been widely used as a biomarker for proliferation in many types of cancer. However, the molecular regulation underlying MCM2 in lung cancer cells is poorly understood. In this study, we investigated the role of MCM2 in lung adenocarcinoma A549 (wild-type p53) and H1299 (null p53) cells. MCM2 overexpression increased cell proliferation in A549 cells while silencing MCM2 decreased cell proliferation in H1299 cells. We performed global quantitative phosphoproteomic analysis to uncover the important downstream networks regulated by MCM2 in lung cancer cells. We identified 1484 phosphorylation sites in 593 phosphoproteins of MCM2-overexpressed A549 cells. Of these phosphosites, 110 phosphoproteins were significantly changed in response to MCM2 overexpression. In addition, we identified 1599 phosphorylation sites in 592 phosphoproteins of MCM2-silenced H1299 cells. Of these phosphosites, 57 phosphoproteins were significantly changed in response to MCM2 silencing. The differentially regulated phosphoproteins are involved in biological functions such as RNA splicing, cell cycle and cytoskeleton regulation. Functional study demonstrated that MCM2 overexpression promoted cell migration in A549 cells. Moreover, silencing MCM2 inhibits cell migration and induces cell cycle arrest in H1299 cells. Furthermore, we observed a common phosphorylation change at Ser-99 of high mobility group protein HMG-I/HMG-Y (HMGA1) in both MCM2 overexpression and silencing, indicating an important regulatory effect of Ser-99 HMGA1 on lung cancer cells. The phosphoproteomic profiling of MCM2 in lung cancer cells provides new insight about phosphorylation networks regulated by MCM2 and reveals novel targets for lung cancer therapy.

Dataset Information

Cell cycle phosphorylation dynamics in vivo reveals switch-like control of intrinsic disorder

Publications

A cyclin-dependent kinase-mediated phosphorylation switch of disordered protein condensation.

Similar Datasets

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