GO:0010389 regulation of G2/M transition of mitotic cell cycle: Cell Cycle Checkpoint Control, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0010389 describes any signaling pathway that modulates the activity of a cyclin-dependent protein kinase to control the switch from G2 phase to M phase of the mitotic cell cycle.
• The core molecular event is activation of the CDK1-cyclin B complex, which is restrained by WEE1/MYT1 kinases and activated by CDC25 phosphatases.
• Checkpoint kinases ATM/ATR and their effectors CHK1/CHK2 delay G2/M transition after DNA damage, providing time for repair.
• The G2/M transition is coordinated with centrosome maturation, Golgi ribbon disassembly, and nuclear mechanobiology.
• Deregulation of G2/M transition is a hallmark of cancer, including melanoma and lung cancer, and is a target for synthetic lethality approaches.
• CRISPR-based models (knockout, point mutation, knock-in, overexpression) enable causal dissection of G2/M regulatory genes in disease contexts.
Description
The G2/M transition is the decisive cell cycle checkpoint where a cell commits to mitosis after completing DNA replication and repairing damage. GO:0010389, regulation of G2/M transition of mitotic cell cycle, captures the signaling pathways that modulate the activity of cyclin-dependent protein kinases to control this switch. This process ensures genomic integrity by integrating inputs from DNA damage sensors, mitotic kinases, and structural remodeling of organelles such as centrosomes and the Golgi apparatus. Researchers study GO:0010389 because its dysregulation is causally linked to cancer progression, and because it offers therapeutic opportunities for synthetic lethality in tumors with defective G2/M checkpoints. The term encompasses both the core CDK1-cyclin B regulatory circuit and the upstream signaling that fine-tunes its timing.
regulation of G2/M transition of mitotic cell cycle At A Glance
| GO ID | GO:0010389 |
|---|---|
| GO term | regulation of G2/M transition of mitotic cell cycle |
| Ontology | biological_process |
| Synonym | regulation of mitotic entry |
| Major function | Modulates CDK1-cyclin B activity to control entry into mitosis |
| Key kinases | CDK1, WEE1, MYT1, CHK1, CHK2, AURKA, PLK1 |
| Key phosphatases | CDC25A, CDC25B, CDC25C |
| Checkpoint sensors | ATM, ATR, and downstream effectors |
| Associated processes | Centrosome maturation, Golgi disassembly, nuclear mechanotransduction |
What Is GO:0010389?
GO:0010389 is defined as any signaling pathway that modulates the activity of a cell cycle cyclin-dependent protein kinase to modulate the switch from G2 phase to M phase of the mitotic cell cycle. In practice, this includes the kinase and phosphatase cascades that activate or inhibit CDK1-cyclin B, the checkpoint kinases that delay mitotic entry in response to DNA damage, and the structural and mechanical cues that coordinate nuclear and organelle remodeling with mitotic commitment.
Why Is regulation of G2/M transition of mitotic cell cycle Important in Cell Biology?
Regulation of the G2/M transition is essential for maintaining genomic stability, because it prevents cells with damaged or incompletely replicated DNA from entering mitosis. The process is also a central node in cancer biology: many tumors override the G2/M checkpoint to proliferate despite genomic stress, and this dependency can be exploited therapeutically through synthetic lethality. Beyond cancer, the G2/M transition is coupled to structural remodeling of the nucleus, centrosomes, and Golgi apparatus, making it a model system for studying how mechanical and organelle-level changes are coordinated with cell cycle progression. Understanding GO:0010389 therefore informs both basic cell biology and the development of targeted anticancer strategies.
• Maintains genomic integrity by delaying mitosis until DNA damage is repaired.
• Controls the activation of CDK1-cyclin B, the master regulator of mitotic entry.
• Integrates DNA damage signals from ATM/ATR and CHK1/CHK2 into cell cycle decisions.
• Coordinates centrosome maturation and mitotic spindle assembly.
• Regulates Golgi ribbon disassembly during mitosis.
• Responds to nuclear mechanobiology and shape changes.
• Is frequently deregulated in melanoma and lung cancer.
• Provides targets for synthetic lethal therapies in checkpoint-defective tumors.
• Involves post-translational modifications such as ubiquitination and mono-ADP-ribosylation.
• Serves as a paradigm for studying cell cycle-dependent organelle remodeling.
What Happens During regulation of G2/M transition of mitotic cell cycle?
Activation of the CDK1-cyclin B complex
In simple terms: The cell's main mitosis engine is turned on.
The core event in G2/M transition is the activation of CDK1 by its regulatory partner cyclin B. CDK1 activity is kept low during G2 by inhibitory phosphorylation at Thr14 and Tyr15, catalyzed by WEE1 and MYT1 kinases. At the G2/M boundary, CDC25 phosphatases remove these inhibitory phosphates, triggering CDK1 activation and commitment to mitosis. This switch is tightly regulated by signaling pathways that modulate the balance between WEE1/MYT1 and CDC25 activities.
DNA damage checkpoint signaling
In simple terms: If DNA is broken, the cell pauses before dividing.
DNA damage activates ATM and ATR kinases, which phosphorylate and activate CHK1 and CHK2. These checkpoint kinases inhibit CDC25 phosphatases and stabilize WEE1, thereby keeping CDK1 inactive and delaying G2/M transition until repair is complete. This checkpoint is critical for preventing the propagation of mutations and is often compromised in cancer cells.
Centrosome maturation and spindle assembly
In simple terms: The cell's skeleton prepares to divide chromosomes.
As cells approach mitosis, centrosomes undergo maturation, a process coordinated with G2/M transition. A cell cycle-dependent transition from acetylation to phosphorylation regulates timely centrosome maturation, ensuring proper spindle formation. Aurora-A kinase, a key regulator of centrosome function, is itself controlled by mono-ADP-ribosylation mediated by PARP10 during G2/M transition.
Golgi ribbon disassembly and nuclear remodeling
In simple terms: The cell's internal compartments reorganize for division.
The Golgi apparatus undergoes ribbon disassembly during G2/M transition, a process that can be studied in vitro and is essential for equal partitioning of organelles. Concurrently, the nucleus experiences mechanical changes, and mechanobiology of the nucleus during G2-M transition influences cell cycle progression. These structural events are integrated with CDK1 activation and checkpoint signaling.
Post-translational regulation of mitotic regulators
In simple terms: Proteins are tagged for destruction or modification to time mitosis.
The G2/M transition is fine-tuned by ubiquitination and other post-translational modifications. For example, TRAP1 controls G2-M transition through regulation of CDK1 and MAD2 expression and ubiquitination. Similarly, alternative splicing of CDC25C, regulated by PUF60, affects cell cycle progression and lung cancer progression. These layers of regulation ensure precise timing and coordination of mitotic entry.
Key Genes Involved in GO:0010389 regulation of G2/M transition of mitotic cell cycle
The following genes and proteins are central to the regulation of G2/M transition, as supported by the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CDK1 | Catalytic subunit of the CDK1-cyclin B complex; drives mitotic entry | Core target for cell cycle inhibition and checkpoint studies |
| CCNB1 | Regulatory cyclin that activates CDK1 | Marker of G2/M progression and target for knockout studies |
| CDC25C | Phosphatase that removes inhibitory phosphates from CDK1 | Regulated by alternative splicing; linked to lung cancer |
| WEE1 | Kinase that phosphorylates and inhibits CDK1 | Target for synthetic lethality in cancer |
| MYT1 | Kinase that inhibits CDK1 at Thr14/Tyr15 | Component of the G2/M checkpoint |
| CHK1 | Effector kinase of ATR; inhibits CDC25 and delays G2/M | Checkpoint regulator and drug target |
| CHK2 | Effector kinase of ATM; inhibits CDC25 | DNA damage response mediator |
| ATM | Sensor kinase activated by DNA double-strand breaks | Upstream regulator of G2/M checkpoint |
| ATR | Sensor kinase activated by replication stress | Upstream regulator of G2/M checkpoint |
| AURKA | Aurora-A kinase; regulates centrosome maturation and mitotic entry | Target of mono-ADP-ribosylation by PARP10 |
| PARP10 | Mono-ADP-ribosylates Aurora-A to regulate G2/M transition | Novel regulator of mitosis |
| TRAP1 | Controls CDK1 and MAD2 expression/ubiquitination | Mitochondrial chaperone linked to G2/M control |
| MAD2 | Spindle assembly checkpoint protein; regulated by TRAP1 | Mitotic checkpoint component |
| PUF60 | Regulates alternative splicing of CDC25C | Promotes cell cycle and lung cancer progression |
| PLK1 | Polo-like kinase 1; activates CDC25 and promotes mitotic entry | Therapeutic target in melanoma |
| CCNB2 | Cyclin B2; partner of CDK1 in mitosis | G2/M progression marker |
| TP53 | Tumor suppressor; induces p21 and G2/M arrest after DNA damage | Frequently mutated in melanoma |
| CDKN1A | p21; inhibits CDK1 and contributes to G2/M checkpoint | Effector of p53-mediated arrest |
How Is regulation of G2/M transition of mitotic cell cycle Regulated?
The regulation of G2/M transition is controlled by multiple signaling inputs. DNA damage activates ATM/ATR-CHK1/CHK2 pathways that inhibit CDC25 and sustain WEE1 activity, delaying mitosis. The PI3K/AKT pathway can modulate WEE1 and CDC25, influencing checkpoint strength. Post-translational modifications, including ubiquitination and mono-ADP-ribosylation, regulate the stability and activity of key mitotic regulators such as CDK1, MAD2, and Aurora-A. Alternative splicing of CDC25C by PUF60 adds another layer of control. Additionally, mechanical cues from the nucleus and organelle remodeling feed into the timing of G2/M transition.
regulation of G2/M transition of mitotic cell cycle and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CDC25C | Lung cancer progression via alternative splicing | Knockout or point mutation in lung cancer cell lines |
| TRAP1 | Cancer cell cycle dysregulation | Knockout and overexpression in tumor models |
| AURKA | Cancer and centrosome amplification | Point mutation of mono-ADP-ribosylation sites |
| CHK1 | Synthetic lethality in melanoma | Knockout in melanoma cell lines |
| PUF60 | Lung cancer | Knockdown or knockout to assess splicing changes |
Cancer
Deregulation of G2/M transition is a hallmark of cancer. In melanoma, targeting the G2-M checkpoint via synthetic lethality is an active therapeutic strategy, especially in tumors with defective DNA damage responses. PUF60 promotes cell cycle progression and lung cancer progression by regulating alternative splicing of CDC25C, highlighting the importance of splicing control in G2/M regulation. TRAP1 controls G2-M transition through CDK1 and MAD2, and its dysregulation may contribute to tumorigenesis.
Melanoma
Melanoma cells frequently exhibit altered cell cycle regulation, including G2/M checkpoint defects. Targeting cell cycle regulation via the G2-M checkpoint for synthetic lethality has been proposed as a therapeutic approach in melanoma. Cell cycle regulation and melanoma reviews emphasize the role of G2/M transition in disease progression and drug resistance.
Lung cancer
PUF60 promotes lung cancer progression by regulating alternative splicing of CDC25C, thereby influencing G2/M transition. This links splicing factors to cell cycle control in lung cancer, suggesting that targeting G2/M regulatory pathways may be beneficial.
Other diseases
While the primary focus is cancer, the G2/M transition is also relevant to developmental disorders and neurodegeneration, though direct evidence from the verified citations is limited to cancer and cell cycle biology. Nuclear mechanobiology during G2-M transition may have implications for diseases involving nuclear envelope defects.
From regulation of G2/M transition of mitotic cell cycle-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of CDK1 abolish G2/M transition? | CDK1 knockout cell lines |
| Does a specific phosphorylation site on CDC25C regulate its activity? | Point mutation knock-in of CDC25C |
| How does mono-ADP-ribosylation of Aurora-A affect G2/M? | Knock-in of PARP10 or Aurora-A mutants |
| What is the effect of TRAP1 overexpression on CDK1 ubiquitination? | Overexpression of TRAP1 in cancer cells |
| Can synthetic lethality be induced by CHK1 inhibition in melanoma? | CHK1 knockout or inhibitor-treated melanoma models |
| How does nuclear mechanobiology influence G2/M transition? | Microfluidic or substrate stiffness models with tagged knock-in of nuclear markers |
How to Study the regulation of G2/M transition of mitotic cell cycle Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Flow cytometry | DNA content and mitotic index | Assessing G2/M arrest and checkpoint function |
| Western blot | Protein expression and phosphorylation | CDK1 activity status |
| Immunoprecipitation | Protein interactions and ubiquitination | TRAP1-mediated CDK1 ubiquitination |
| Fluorescence microscopy | Organelle and centrosome dynamics | Golgi disassembly and centrosome maturation |
| CRISPR knockout screening | Gene essentiality and synthetic lethality | Identifying G2/M regulators in cancer |
| RNA-seq | Transcriptional changes and splicing | Alternative splicing of CDC25C |
| Proteomics | Global protein expression and modifications | Mono-ADP-ribosylation of Aurora-A |
| Live-cell imaging | Real-time mitotic progression | Nuclear mechanobiology during G2-M |
Cell cycle synchronization and flow cytometry
To study G2/M transition, cells are synchronized using drugs such as nocodazole or thymidine block, and progression is monitored by flow cytometry for DNA content and mitotic markers. This method quantifies the proportion of cells in G2/M and assesses checkpoint function.
Western blotting and immunoprecipitation
Western blotting detects phosphorylation states of CDK1 (Thr14/Tyr15), CDC25C, and other regulators, while immunoprecipitation can assess ubiquitination and protein interactions. These techniques are essential for mechanistic studies of G2/M regulation.
Imaging of organelles and centrosomes
Fluorescence microscopy visualizes centrosome maturation, Golgi ribbon disassembly, and nuclear morphology during G2/M transition. Live-cell imaging with tagged proteins allows dynamic tracking of mitotic entry.
CRISPR screening and functional genomics
Genome-wide CRISPR knockout screens can identify genes that regulate G2/M transition or confer sensitivity to checkpoint inhibitors. These screens are powerful for discovering synthetic lethal interactions.
How CRISPR Can Be Used to Study GO:0010389 regulation of G2/M transition of mitotic cell cycle
Knockout
CRISPR knockout of genes such as CDK1, CDC25C, or CHK1 can abolish or delay G2/M transition, providing causal evidence for their roles. Knockout cell lines are also used to test synthetic lethality with checkpoint inhibitors.
Point Mutation
Point mutations can be introduced to study specific phosphorylation or modification sites, such as the inhibitory phosphorylation sites on CDK1 or the mono-ADP-ribosylation site on Aurora-A. These models help dissect precise molecular mechanisms.
Knock-in
Knock-in of tagged proteins (e.g., GFP-CDK1 or luciferase reporters) allows real-time monitoring of G2/M transition in live cells. Knock-in of disease-associated mutations can model cancer-related dysregulation.
Overexpression
Overexpression of regulators like TRAP1 or PUF60 can drive cell cycle progression and tumorigenesis, mimicking gain-of-function events in cancer. These models are useful for testing targeted therapies.
How EDITGENE Supports regulation of G2/M transition of mitotic cell cycle Research
Researchers studying regulation of G2/M transition of mitotic cell cycle-related genes often need to determine whether a candidate gene is causally involved in mitotic entry, checkpoint control, or disease progression. EDITGENE provides a comprehensive suite of CRISPR-based services to generate precisely engineered cell models, enabling rigorous functional validation of G2/M regulatory genes.
Contact EDITGENE today to design your custom CRISPR model for regulation of G2/M transition of mitotic cell cycle research.
Frequently Asked Questions About regulation of G2/M transition of mitotic cell cycle
What is GO:0010389?
GO:0010389 is the Gene Ontology term for regulation of G2/M transition of mitotic cell cycle, describing signaling pathways that modulate CDK1 activity to control entry into mitosis.
What genes are involved in regulation of G2/M transition?
Key genes include CDK1, CCNB1, CDC25C, WEE1, MYT1, CHK1, CHK2, ATM, ATR, AURKA, PARP10, TRAP1, and PUF60.
How is the G2/M transition regulated?
It is regulated by the balance between inhibitory kinases (WEE1/MYT1) and activating phosphatases (CDC25), as well as checkpoint kinases that delay mitosis after DNA damage.
Why is the G2/M checkpoint important in cancer?
Cancer cells often have defective G2/M checkpoints, making them dependent on remaining checkpoint proteins for survival, which can be exploited by synthetic lethality.
What methods are used to study G2/M transition?
Common methods include flow cytometry, western blotting, immunofluorescence, CRISPR screening, and live-cell imaging.
What is the role of CDK1 in G2/M transition?
CDK1 is the catalytic subunit that, when activated by cyclin B and dephosphorylated by CDC25, drives cells into mitosis.
How does DNA damage delay G2/M transition?
DNA damage activates ATM/ATR, which through CHK1/CHK2 inhibit CDC25 and stabilize WEE1, keeping CDK1 inactive and delaying mitosis.
What is the connection between G2/M transition and centrosomes?
Centrosome maturation is coordinated with G2/M transition, and Aurora-A kinase regulates this process.
Can CRISPR be used to study G2/M transition?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect gene function in G2/M transition.
What diseases are linked to G2/M transition dysregulation?
Cancer, particularly melanoma and lung cancer, is strongly linked to dysregulation of G2/M transition.
Conclusion
GO:0010389, regulation of G2/M transition of mitotic cell cycle, is a fundamental biological process that ensures accurate mitotic entry and genomic stability. Its core machinery, centered on CDK1-cyclin B and checkpoint kinases, is finely tuned by phosphorylation, ubiquitination, and splicing. Dysregulation of this process contributes to cancer, making it a prime target for therapeutic intervention. Advances in CRISPR-based models and functional genomics continue to illuminate new regulators and disease connections, offering hope for precision medicine approaches.
References
- 1. Xu N et al.. 2023. PUF60 promotes cell cycle and lung cancer progression by regulating alternative splicing of CDC25C.. Cell Rep 42(9):113041 PMID: 37682709
- 2. Lima JT et al.. 2024. Mechanobiology of the nucleus during the G2-M transition.. Nucleus 15(1):2330947 PMID: 38533923
- 3. Sisinni L et al.. 2017. TRAP1 controls cell cycle G2-M transition through the regulation of CDK1 and MAD2 expression/ubiquitination.. J Pathol 243(1):123-134 PMID: 28678347
- 4. Di Paola S et al.. 2022. PARP10 Mediates Mono-ADP-Ribosylation of Aurora-A Regulating G2/M Transition of the Cell Cycle.. Cancers (Basel) 14(21) PMID: 36358629
- 5. Barnaba N et al.. 2021. Targeting cell cycle regulation via the G2-M checkpoint for synthetic lethality in melanoma.. Cell Cycle 20(11):1041-1051 PMID: 33966611
- 6. Li J et al.. 2026. A cell cycle-dependent transition of acetylation to phosphorylation regulates timely centrosome maturation.. Nat Commun 17(1) PMID: 41862458
- 7. Xu W et al.. 2016. Cell Cycle Regulation and Melanoma.. Curr Oncol Rep 18(6):34 PMID: 27106898
- 8. Ayala I et al.. 2023. In Vitro Methods to Investigate the Disassembly of the Golgi Ribbon During the G2-M Transition of the Cell Cycle.. Methods Mol Biol 2557:333-347 PMID: 36512225
