Positively charged specificity site in cyclin B1 is essential for mitotic fidelity DOI Creative Commons
Thomas U. Mayer,

Christian Heinzle,

Anna Höfler

et al.

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: April 9, 2024

Abstract Phosphorylation of substrates by cyclin-dependent kinases (CDKs) is the driving force cell cycle progression. Several CDK-activating cyclins are involved, yet how they contribute to substrate specificity still poorly understood. Here, we discovered that a positively charged pocket in cyclin B1, which exclusively conserved within B-type and binds phosphorylated serine- or threonine-residues, essential for correct execution mitosis. HeLa cells expressing mutant B1 strongly delayed anaphase onset due multiple defects mitotic spindle function timely activation E3 ligase APC/C. Pocket integrity APC/C phosphorylation particularly at non-consensus CDK1 sites full vitro ubiquitylation activity. Our results support model B1’s serves as site factor sequential phosphorylations involving initial priming events facilitate subsequent pocket-dependent even motifs.

Language: Английский

Positively charged specificity site in cyclin B1 is essential for mitotic fidelity DOI Creative Commons

Christian Heinzle,

Anna Höfler, Jun Yu

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 20, 2025

Abstract Phosphorylation of substrates by cyclin-dependent kinases (CDKs) is the driving force cell cycle progression. Several CDK-activating cyclins are involved, yet how they contribute to substrate specificity still poorly understood. Here, we discover that a positively charged pocket in cyclin B1, which exclusively conserved within B-type and binds phosphorylated serine- or threonine-residues, essential for correct execution mitosis. HeLa cells expressing mutant B1 strongly delayed anaphase onset due multiple defects mitotic spindle function timely activation E3 ligase APC/C. Pocket integrity APC/C phosphorylation particularly at non-consensus CDK1 sites full vitro ubiquitylation activity. Our results support model B1’s facilitates sequential phosphorylations involving initial priming events assist subsequent pocket-dependent even motifs.

Language: Английский

Citations

1
Phosphate-binding pocket on cyclin B governs CDK substrate phosphorylation and mitotic timing DOI Creative Commons
Henry Ng, Armin N. Adly, Devon H. Whelpley

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 29, 2024

Summary Cell cycle progression is governed by complexes of the cyclin-dependent kinases (CDKs) and their regulatory subunits cyclin Cks1. CDKs phosphorylate hundreds substrates, often at multiple sites. Multisite phosphorylation depends on Cks1, which binds initial priming sites to promote secondary other Here, we describe a similar role for recently discovered phosphate-binding pocket (PP) B-type cyclins. Mutation PP in Clb2, major mitotic budding yeast, alters bud morphology delays onset anaphase. Using phosphoproteomics vivo kinase reactions vitro , find that mutation reduces several CDK including Bud6 subunit polarisome Cdc16 Cdc27 anaphase-promoting complex/cyclosome. We conclude PP, like controls timing multisite thereby helping establish robust cell-cycle events.

Language: Английский

Citations

4
The Yeast Gsk-3 Kinase Mck1 Is Necessary for Cell Wall Remodeling in Glucose-Starved and Cell Wall-Stressed Cells DOI Open Access
Fan Zhang,

Yingzhi Tang,

Houjiang Zhou

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(8), P. 3534 - 3534

Published: April 9, 2025

The cell wall integrity (CWI) pathway is responsible for transcriptional regulation of remodeling in response to stress. How mediated by the CWI effected inputs from other signaling pathways not well understood. Here, we demonstrate that Mck1 kinase cooperates with Slt2, MAP pathway, promote thickening glucose-starved cells. Integrative analyses transcriptome, proteome and metabolic profiling indicate required accumulation UDP-glucose (UDPG), substrate β-glucan synthesis, through activation two regulons: Msn2/4-dependent stress Cat8-/Adr1-mediated reprogram dependent on SNF1 complex. Analysis phosphoproteome suggests similar mammalian Gsk-3 kinases, involved cytoskeleton-dependent cellular processes, metabolism, transcription. Specifically, may be implicated Snf1-dependent PKA inhibition SAGA (Spt-Ada-Gcn5 acetyltransferase)-mediated transcription activation, a hypothesis further underscored significant overlap between Mck1- Gcn5-activated transcriptomes. Phenotypic analysis also supports roles actin cytoskeleton-mediated exocytosis ensure plasma membrane homeostasis wall-stressed Together, these findings only reveal novel functions reprogramming polarized growth but provide valuable omics resources future studies uncover underlying mechanisms kinases response.

Language: Английский

Citations

0
Phosphate-binding pocket on cyclin B governs CDK substrate phosphorylation and mitotic timing DOI Creative Commons
Henry Ng, Devon H. Whelpley, Armin N. Adly

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: May 8, 2025

Abstract Cell cycle progression is governed by complexes of the cyclin-dependent kinases (CDKs) and their regulatory subunits cyclin Cks1. CDKs phosphorylate hundreds substrates, often at multiple sites. Multisite phosphorylation depends on Cks1, which binds initial priming sites to promote secondary other Here, we describe a similar role for recently discovered phosphate-binding pocket (PP) B-type cyclins. Mutation PP in Clb2, major mitotic budding yeast, alters bud morphology delays onset anaphase. reduces multi-site CDK substrates vitro, including Cdc16 Cdc27 anaphase-promoting complex/cyclosome Bud6 Spa2 polarisome. We conclude that PP, like controls pattern multisite thereby helping establish robust timing cell-cycle events.

Language: Английский

Citations

0
Whi5 hypo- and hyper-phosphorylation dynamics control cell-cycle entry and progression DOI
Jordan Xiao, Jonathan J. Turner, Mardo Kõivomägi

et al.

Current Biology, Journal Year: 2024, Volume and Issue: 34(11), P. 2434 - 2447.e5

Published: May 14, 2024

Language: Английский

Citations

3
Crystallographic fragment screening of CDK2-cyclin A: FragLites map sites of protein-protein interaction DOI Creative Commons
Ian Hope,

Martin E. M. Noble,

Michael J. Waring

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 3, 2024

Abstract Protein-protein interaction sites (PPIs) are potentially more selective therapeutic binding than protein substrate sites. PPIs include distinct regions frequently called “hotspots,” of key amino acid interactions. Prospective identification these hotspots through X-ray crystallographic screening could assist in the separation function mutants for experimental validation, enhance confidence AI-generated multiprotein complex predictions and accelerate development chemical probes. To explore applications, we utilize FragLite library to examine surfaces CDK2-cyclin A. The many protein- peptide-CDK2-cyclin A complexes that have been structurally characterised make this an appropriate test case. We show FragLites comprehensively map both known protein-protein on identify a possible uncharacterised site, providing structural method toward directing mechanistic studies starting points probe design.

Language: Английский

Citations

2
The substrate quality of CK2 target sites has a determinant role on their function and evolution DOI Creative Commons
David Bradley, Chantal Garand, Hugo Belda

et al.

Cell Systems, Journal Year: 2024, Volume and Issue: 15(6), P. 544 - 562.e8

Published: June 1, 2024

Language: Английский

Citations

2
Phosphorylation of Orc6 During Mitosis Regulates DNA Replication and Ribosome Biogenesis DOI
Fredy Kurniawan, Arindam Chakraborty,

Humayra Z. Oishi

et al.

Molecular and Cellular Biology, Journal Year: 2024, Volume and Issue: 44(7), P. 289 - 301

Published: June 12, 2024

The human Origin Recognition Complex (ORC) is required not only for the initiation of DNA replication, but also implicated in diverse cellular functions, including chromatin organization, centrosome biology, and cytokinesis. smallest subunit ORC, Orc6, poorly conserved amongst eukaryotes. Recent studies from our laboratory have suggested that Orc6 replication licensing, needed S-phase progression. Further, ATR-dependent phosphorylation at T229 damage response during S-phase. In this study, we demonstrate CDK-dependent T195 occurs mitosis. While does seem to be exit mitosis, cells expressing phosphomimetic T195E mutant impede Moreover, phosphorylated form associates with ORC more robustly, shows enhanced association outside G1, supporting view may prevent role Orc1-5 licensing G1. Finally, localize nucleolar organizing centers regulate ribosome biogenesis. Our results suggest prevents replication.

Language: Английский

Citations

2
Cyclin B3 is a dominant fast-acting cyclin that drives rapid early embryonic mitoses DOI
Pablo Lara-González,

Smriti Variyar,

Shabnam Moghareh

et al.

The Journal of Cell Biology, Journal Year: 2024, Volume and Issue: 223(11)

Published: Aug. 6, 2024

Mitosis in early embryos often proceeds at a rapid pace, but how this pace is achieved not understood. Here, we show that cyclin B3 the dominant driver of embryonic mitoses C. elegans embryo. Cyclins B1 and B2 support slow mitosis (NEBD to anaphase ∼600 s), presence dominantly drives approximately threefold faster observed wildtype. Multiple mitotic events are slowed down B2-driven mitosis, B3-associated Cdk1 H1 kinase activity ∼25-fold more active than B1-associated Cdk1. Addition fast B3-only introduces an ∼60-s delay between completion chromosome alignment onset; delay, which important for segregation fidelity, dependent on inhibitory phosphorylation activator Cdc20. Thus, dominance, coupled B1-dependent acts via Cdc20 phosphorylation, sets ensures fidelity

Language: Английский

Citations

2
The role of intrinsic protein disorder in regulation of cyclin-dependent kinases DOI
Aaron H. Phillips, Richard W. Kriwacki

Current Opinion in Structural Biology, Journal Year: 2024, Volume and Issue: 88, P. 102906 - 102906

Published: Aug. 13, 2024

Language: Английский

Citations

2