Summary of ssbd-repos-000325

Name
URL
DOI

Title
Dataset for "Live-cell imaging provides direct evidence for a threshold in CDK activity at the G2/M transition."
Description

Cyclin-dependent kinase (CDK) plays an essential role in determining the temporal ordering of the cell cycle phases. However, despite significant progress in studying regulators of CDK and phosphorylation patterns of CDK substrates at the population level, it remains elusive how CDK regulators coordinately affect CDK activity at the single-cell level and how CDK controls the temporal order of cell cycle events. This is mostly due to the lack of tools for quantifying CDK activity in individual living cells. Here, we elucidate the dynamics of CDK activity in fission yeast and mammalian cells by using a newly developed CDK activity biosensor, Eevee-spCDK, based on Förster Resonance Energy Transfer (FRET). Taking advantage of this system, we unravel the profile of CDK activity in vegetatively growing S. pombe cells. Thus, we detect a transient increase in S phase followed by a gradual increment during G2 phase. CDK activity then reaches its maximum in early M phase and rapidly decreases at mitotic exit. During G2 phase, CDK activity exhibits a biphasic pattern, i.e. an early slow increase and a late fast rise prior to the G2/M phase transition, as predicted from mathematical studies. Remarkably, although CDK activity does not necessarily correlate with cyclin levels, we find that it converges to the same level around mitotic onset in several mutant backgrounds, including pom1Δ cells and wee1 or cdc25 overexpressing cells. These data provide the first direct evidence that cells enter M phase when CDK activity reaches a high threshold, consistent with the quantitative model of cell cycle progression in fission yeast.

Submited Date
2023-09-27
Release Date
2024-01-15
Updated Date
-
License
Funding information
-
File formats
Tiff
Data size
76.2 GB

Organism
Schizosaccharomyces pombe
Strain
See details in Sugiyama et. al., bioRxiv. 2023
Cell Line
Hela cell
Genes
-
Proteins
-

GO Molecular Function (MF)
Cyclin-dependent kinase (CDK)
GO Biological Process (BP)
Cell cycle regulation
GO Cellular Component (CC)
Cell
Study Type
Cyclin-dependent kinase (CDK)
Imaging Methods
spinning disk confocal microscopy, wide-field microscopy

Method Summary

See details in Sugiyama et. al., bioRxiv. 2023

Related paper(s)

Hironori Sugiyama, Yuhei Goto, Yohei Kondo, Damien Coudreuse, Kazuhiro Aoki (2023/01/01), Live-cell imaging provides direct evidence for a threshold in CDK activity at the G2/M transition, bioRxiv, 2023.03.26.534249

Published in 2023/01/01

(Abstract) Cyclin-dependent kinase (CDK) plays an essential role in determining the temporal ordering of the cell cycle phases. However, despite significant progress in studying regulators of CDK, it remains elusive how they coordinately affect CDK activity at the single-cell level and how CDK controls the temporal order of cell cycle events. This could be due to the lack of tools to monitor CDK activity in living cells. Here, we elucidate the dynamics of CDK activity in fission yeast and mammalian cells by using a newly developed CDK activity biosensor, Eevee-spCDK, based on Förster Resonance Energy Transfer (FRET). Taking advantage of this system, we unravel the profile of CDK activity in vegetatively growing S. pombe cells. Thus, we detect a transient increase in S phase followed by a gradual increment during G2 phase. CDK activity then reaches its maximum in early M phase and rapidly decreases at mitotic exit. During G2 phase, CDK activity exhibits a biphasic pattern, i.e., an early slow increase and a late fast rise prior to the G2/M phase transition, as predicted from mathematical studies. Remarkably, although CDK activity does not necessarily correlate with cyclin levels, we find that it converges to the same level around mitotic onset in several mutant backgrounds, including pom1Δ cells and wee1 or cdc25 overexpressing cells. These data provide the first direct evidence that cells enter M phase when CDK activity reaches a high threshold, consistent with the quantitative model of cell cycle progression in fission yeast.Competing Interest StatementThe authors have declared no competing interest.
Related paper(s)

Hironori Sugiyama, Yuhei Goto, Yohei Kondo, Damien Coudreuse, Kazuhiro Aoki (2024) Live-cell imaging defines a threshold in CDK activity at the G2/M transition., Developmental cell

Published in 2024 Jan 8 (Electronic publication in Jan. 8, 2024, midnight )

(Abstract) Cyclin-dependent kinase (CDK) determines the temporal ordering of the cell cycle phases. However, despite significant progress in studying regulators of CDK and phosphorylation patterns of CDK substrates at the population level, it remains elusive how CDK regulators coordinately affect CDK activity at the single-cell level and how CDK controls the temporal order of cell cycle events. Here, we elucidate the dynamics of CDK activity in fission yeast and mammalian cells by developing a CDK activity biosensor, Eevee-spCDK. We find that although CDK activity does not necessarily correlate with cyclin levels, it converges to the same level around mitotic onset in several mutant backgrounds, including pom1Delta cells and wee1 or cdc25 overexpressing cells. These data provide direct evidence that cells enter the M phase when CDK activity reaches a high threshold, consistent with the quantitative model of cell cycle progression in fission yeast.

Contact(s)
Kazuhiro Aoki
Organization(s)
National Institutes of Natural Sciences , National Institute for Basic Biology, Exploratory Research Center for Life and Living Systems , Division of Quantitative Biology
Image Data Contributors
Hironori Sugiyama, Yuhei Goto
Quantitative Data Contributors
Hironori Sugiyama, Yuhei Goto

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