GO:0000281 mitotic cytokinesis: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0000281 mitotic cytokinesis is the cell cycle process that divides the cytoplasm after mitosis, separating one cell into two daughter cells.
• Mitotic exit and cytokinesis are spatially and temporally coordinated by the APC/C, PP1/PP2A phosphatases and mitotic kinases such as Plk1.
• The Dma1-mediated mitotic checkpoint couples chromosome segregation to cytokinesis, preventing premature abscission.
• Animal cytokinesis requires symmetry-breaking cues that position and constrict the actomyosin ring.
• Lipid polarization at the cleavage furrow contributes to membrane remodeling during cytokinesis.
• Errors in cytokinesis are linked to chromosomal instability and oncogenesis, making these genes attractive cancer research targets.
Description
Mitotic cytokinesis (GO:0000281) is the final step of the cell cycle, in which the cytoplasm of a mitotic cell is divided to produce two separate daughter cells. This process is not a passive consequence of mitosis but an actively regulated program that must be coordinated with chromosome segregation, mitotic exit and membrane remodeling. Because cytokinesis failure generates binucleated or tetraploid cells, its regulation is central to genome stability and tumor suppression. Researchers study mitotic cytokinesis to understand how cells spatially and temporally couple the end of mitosis to physical division, and how disruption of this coupling contributes to disease. The process is driven by conserved machinery including the actomyosin contractile ring, the centrosome, and regulatory kinases and phosphatases that establish the site and timing of furrow ingression. In recent years, quantitative imaging, genetics and proteomics in model organisms such as fission yeast and mammalian cells have revealed that cytokinesis nodes scale with cell size and that lipid polarization contributes to membrane dynamics at the cleavage furrow. These findings make GO:0000281 a rich ontology term for both fundamental cell biology and translational cancer research.
mitotic cytokinesis At A Glance
| GO ID | GO:0000281 |
|---|---|
| GO term | mitotic cytokinesis |
| Ontology | biological_process |
| Synonym | cytokinesis after mitosis |
| Major function | Division of the cytoplasm after mitosis to separate one cell into two daughter cells |
| Process context | Cell cycle process occurring at the end of mitosis |
| Key regulators | APC/C, PP1, PP2A, Plk1, Dma1 checkpoint |
| Model systems | Fission yeast, mammalian cultured cells, animal models |
What Is GO:0000281?
In our own words, GO:0000281 mitotic cytokinesis is the biological process that divides the cytoplasm of a cell after mitosis, resulting in the separation of the original cell into two daughter cells. It is a cell cycle process that follows nuclear division and encompasses the assembly and constriction of the division machinery, membrane remodeling and final abscission. The QuickGO synonym cytokinesis after mitosis captures its placement at the end of the mitotic program.
Why Is mitotic cytokinesis Important in Cell Biology?
Mitotic cytokinesis is essential because it physically completes cell division and safeguards genome stability; failure of this process produces binucleated cells and aneuploidy, which are hallmarks of chromosomal instability and cancer. The process is also a paradigm for understanding how cells integrate spatial cues, kinase and phosphatase signaling, and membrane trafficking to execute a mechanical task. Because cytokinesis must be tightly coordinated with mitotic exit, it provides a window into checkpoint control and the consequences of deregulated cell cycle timing.
• Completes cell division by separating the cytoplasm after mitosis.
• Prevents binucleation and tetraploidy, which can drive chromosomal instability.
• Coordinates mitotic exit with physical division through APC/C and phosphatases.
• Requires symmetry-breaking to position the cleavage furrow correctly.
• Involves Plk1 and other mitotic kinases that link mitotic entry to cytokinesis.
• Depends on actomyosin ring assembly and constriction.
• Involves lipid polarization and membrane remodeling at the furrow.
• Is monitored by checkpoints such as the Dma1-mediated mitotic checkpoint.
• Provides targets for cancer research because cytokinesis errors promote oncogenesis.
• Is studied quantitatively in fission yeast where node number scales with cell size.
What Happens During mitotic cytokinesis?
Initiation and symmetry-breaking
In simple terms: The cell first decides exactly where to pinch in two.
After mitosis, the cell must choose a division plane and break symmetry to position the cleavage furrow. Animal cytokinesis relies on symmetry-breaking mechanisms that convert global signals into a localized contractile zone. This step ensures that the furrow forms at the correct position relative to the mitotic spindle and chromosomes.
Assembly of the division machinery
In simple terms: The cell builds a ring that will squeeze the cell in half.
The actomyosin contractile ring assembles at the equatorial cortex and generates the force for furrow ingression. In fission yeast, the number of cytokinesis nodes that template this ring scales with cell size, providing a quantitative link between cell geometry and division machinery. Assembly is coordinated with mitotic exit signals so that ring formation occurs only after chromosome segregation.
Constriction and membrane remodeling
In simple terms: The ring tightens and the membrane follows to pinch the cell.
Constriction of the actomyosin ring drives furrow ingression, while lipid polarization and membrane trafficking deliver new membrane to the growing furrow. Lipid polarization during cytokinesis contributes to the asymmetric distribution of lipids that supports membrane curvature and remodeling. These events are tightly coupled to the mechanical constriction of the ring.
Coordination with mitotic exit
In simple terms: The cell does not divide until it has finished separating its chromosomes.
Mitotic exit and cytokinesis are spatially and temporally controlled by the APC/C and by PP1 and PP2A phosphatases. The APC/C triggers degradation of mitotic cyclins and other substrates, while PP1 and PP2A reverse mitotic phosphorylations to license cytokinesis. The Dma1-mediated mitotic checkpoint further coordinates mitosis with cytokinesis, preventing premature abscission when chromosome segregation is incomplete.
Abscission and completion
In simple terms: The final cut separates the two daughter cells.
The terminal step of cytokinesis, abscission, physically separates the two daughter cells. This step must be carefully timed with mitotic exit and checkpoint signaling to avoid cutting DNA or generating aneuploid cells. Plk1 and other mitotic kinases contribute to the regulation of events from mitotic entry through cytokinesis, including the final stages of division.
Key Genes Involved in GO:0000281 mitotic cytokinesis
The following genes and proteins are central to mitotic cytokinesis and are frequently studied in this context.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ANAPC (APC/C subunits) | Ubiquitin ligase controlling mitotic exit and cytokinesis | Core regulator of the transition from mitosis to cytokinesis |
| PLK1 | Polo-like kinase regulating mitotic entry and cytokinesis | Key mitotic kinase linking mitosis to cytokinesis |
| PPP1CA (PP1) | Protein phosphatase 1 reversing mitotic phosphorylations | Spatial and temporal control of mitotic exit and cytokinesis |
| PPP2CA (PP2A) | Protein phosphatase 2A regulating mitotic exit | Coordinates dephosphorylation events required for cytokinesis |
| DMA1 | Checkpoint E3 ligase coupling mitosis with cytokinesis | Prevents premature cytokinesis when mitosis is incomplete |
| RhoA | Small GTPase promoting actomyosin ring assembly | Central to contractile ring formation and furrow ingression |
| Actin (ACTB/ACT1) | Structural component of the contractile ring | Provides force for cleavage furrow ingression |
| Myosin II (MYH9/MYO2) | Motor protein driving ring constriction | Generates contractile force during cytokinesis |
| Anillin (ANLN) | Scaffold protein organizing the contractile ring | Links actin, myosin and membrane during cytokinesis |
| ECT2 | RhoA guanine nucleotide exchange factor | Activates RhoA at the cleavage furrow |
| Citron kinase (CIT) | RhoA effector regulating contractile ring | Required for cytokinesis in some cell types |
| Formins (e.g. DIAPH1) | Actin nucleators in the contractile ring | Promote actin filament assembly during cytokinesis |
| Septins | Filament-forming proteins at the cleavage furrow | Contribute to membrane and ring organization |
| Cep55 | Centrosomal protein required for abscission | Recruits ESCRT machinery to the midbody |
| ESCRT-III (CHMP4B) | Membrane scission machinery | Executes final abscission step |
| Aurora B (AURKB) | Chromosomal passenger kinase | Regulates cleavage furrow positioning and abscission |
| Cdk1 (CDK1) | Cyclin-dependent kinase driving mitosis | Its inactivation is required for mitotic exit and cytokinesis |
How Is mitotic cytokinesis Regulated?
Mitotic cytokinesis is regulated by the APC/C, which triggers degradation of mitotic cyclins and other substrates to promote mitotic exit. PP1 and PP2A phosphatases provide spatial and temporal control by reversing mitotic phosphorylations, thereby licensing cytokinesis. The Dma1-mediated mitotic checkpoint coordinates mitosis with cytokinesis, ensuring that division does not proceed prematurely. Plk1 and other mitotic kinases also contribute to the regulation of events from mitotic entry through cytokinesis. In addition, lipid polarization and membrane trafficking regulate the membrane remodeling required for furrow ingression and abscission.
mitotic cytokinesis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PLK1 | Cancer, chromosomal instability | Knockout or point-mutation cell lines to test kinase function |
| ANAPC (APC/C subunits) | Cancer, cell cycle deregulation | Knockout models to study mitotic exit defects |
| PPP1CA/PPP2CA | Cell cycle defects, potential cancer relevance | Point-mutation knock-in to dissect phosphatase regulation |
| DMA1 | Genome instability, checkpoint dysfunction | Knockout and tagged knock-in to monitor checkpoint activity |
| RhoA/ECT2 | Cytokinesis failure, potential oncogenesis | Overexpression and knockout to test furrow formation |
Cytokinesis errors and cancer
Errors in mitotic kinases that control duplication, segregation and cytokinesis can lead to chromosomal instability and oncogenesis. Because cytokinesis failure produces binucleated and tetraploid cells, deregulation of this process is mechanistically linked to tumor development. Genes such as PLK1 and APC/C subunits are therefore studied as potential cancer targets.
Checkpoint dysfunction and genome instability
The Dma1-mediated mitotic checkpoint couples mitosis with cytokinesis, and its dysfunction can allow premature abscission or failed division. Such defects contribute to genome instability, a hallmark of many cancers. Studying this checkpoint provides insight into how cells safeguard genome integrity during division.
Phosphatase deregulation in disease
PP1 and PP2A control mitotic exit and cytokinesis, and their deregulation has been implicated in cell cycle defects associated with disease. Because these phosphatases are spatially and temporally controlled, their misregulation can disrupt the coordination between mitosis and cytokinesis.
From mitotic cytokinesis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is a candidate gene required for cytokinesis? | Knockout cell line followed by imaging of furrow ingression |
| Does a specific phosphorylation site regulate cytokinesis timing? | Point-mutation knock-in of the phospho-site |
| Where and when is a protein localized during cytokinesis? | Tagged knock-in with fluorescent protein |
| Does overexpression of a gene drive multinucleation? | Overexpression cell model with nuclei counting |
| How does cell size affect cytokinesis node number? | Quantitative imaging in fission yeast |
| Does a mutation disrupt lipid polarization at the furrow? | Knock-in or knockout with lipid probes |
How to Study the mitotic cytokinesis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Live-cell fluorescence imaging | Dynamics of furrow ingression and abscission | Testing gene requirements for cytokinesis |
| Fixed-cell immunofluorescence | Localization of ring and midbody proteins | Validating tagged knock-in lines |
| Node counting in fission yeast | Number of cytokinesis nodes per cell | Studying scaling with cell size |
| Kinase/phosphatase activity assays | Enzymatic activity of Plk1, PP1, PP2A | Dissecting regulatory phosphorylation |
| Lipid polarization imaging | Distribution of lipids at the cleavage furrow | Membrane remodeling studies |
| Time-lapse microscopy | Timing of mitotic exit and cytokinesis | Checkpoint coordination studies |
| Genetic knockout screens | Identification of cytokinesis genes | Discovery of new regulators |
| Proteomics of midbody | Protein composition of the midbody | Identifying abscission machinery |
Live-cell imaging of cytokinesis
Live-cell imaging with fluorescently tagged actomyosin ring components allows direct visualization of furrow ingression and abscission timing. This method is essential for determining whether a gene perturbation affects the initiation, constriction or completion of cytokinesis.
Quantitative analysis of cytokinesis nodes
In fission yeast, the number of cytokinesis nodes can be counted and correlated with cell size, providing a quantitative readout of division machinery scaling. This approach is useful for testing how genetic perturbations alter node assembly.
Phosphatase and kinase activity assays
Because PP1, PP2A and Plk1 regulate mitotic exit and cytokinesis, kinase and phosphatase activity assays help define the signaling events that control division timing. These assays can be combined with point mutations to test specific regulatory sites.
Lipid polarization imaging
Lipid probes and membrane markers can be used to monitor lipid polarization during cytokinesis and to test how perturbations affect membrane remodeling at the furrow. This method complements protein-based imaging of the contractile ring.
How CRISPR Can Be Used to Study GO:0000281 mitotic cytokinesis
Knockout
CRISPR knockout of candidate cytokinesis genes allows researchers to test whether the gene is required for furrow ingression or abscission. Knockout cell lines can be imaged to quantify binucleation and division failure.
Point Mutation
Point-mutation knock-in can be used to test the function of specific phosphorylation sites in regulators such as PP1, PP2A or Plk1. This approach distinguishes catalytic activity from regulatory site function.
Knock-in
Tagged knock-in of cytokinesis genes with fluorescent proteins enables live tracking of protein localization during division. Knock-in of disease-relevant variants can model how mutations affect cytokinesis.
Overexpression
Overexpression of cytokinesis regulators such as RhoA or ECT2 can drive ectopic furrow formation or multinucleation, providing a gain-of-function readout. Overexpression models are useful for testing whether a gene is sufficient to perturb division.
How EDITGENE Supports mitotic cytokinesis Research
Researchers studying mitotic cytokinesis-related genes often need to determine whether a candidate gene is causally involved in furrow ingression, abscission or checkpoint control. EDITGENE provides CRISPR-based cell models and screening services to support these studies.
Contact EDITGENE today to design your custom CRISPR model for mitotic cytokinesis research.
Frequently Asked Questions About mitotic cytokinesis
What is mitotic cytokinesis (GO:0000281)?
Mitotic cytokinesis is the cell cycle process that divides the cytoplasm after mitosis, separating one cell into two daughter cells.
What genes are involved in mitotic cytokinesis?
Key genes include APC/C subunits, PLK1, PP1, PP2A, DMA1, RhoA, ECT2, anillin, myosin II, septins, Cep55 and ESCRT-III components.
How is mitotic cytokinesis regulated?
It is regulated by the APC/C, PP1 and PP2A phosphatases, Plk1 and the Dma1-mediated mitotic checkpoint, which coordinate mitotic exit with division.
Why is cytokinesis important for cancer research?
Cytokinesis errors cause chromosomal instability and oncogenesis, making cytokinesis genes potential cancer targets.
What happens if cytokinesis fails?
Failure of cytokinesis produces binucleated or tetraploid cells, which can contribute to genome instability.
What is the role of Plk1 in cytokinesis?
Plk1 is a mitotic kinase that regulates events from mitotic entry through cytokinesis, including furrow formation and abscission.
How do PP1 and PP2A control cytokinesis?
They reverse mitotic phosphorylations to provide spatial and temporal control of mitotic exit and cytokinesis.
What is the Dma1-mediated mitotic checkpoint?
It is a checkpoint that couples mitosis with cytokinesis to prevent premature division.
How can I study cytokinesis in the lab?
Live-cell imaging, node counting in fission yeast, kinase/phosphatase assays and lipid polarization imaging are common approaches.
Does cell size affect cytokinesis?
In fission yeast, the number of cytokinesis nodes scales with cell size, linking geometry to division machinery.
Conclusion
Mitotic cytokinesis (GO:0000281) is the essential final step of the cell cycle that divides the cytoplasm after mitosis. It is controlled by a conserved network of kinases, phosphatases and checkpoint proteins that coordinate mitotic exit with physical division. Because cytokinesis failure leads to genome instability and cancer, this process remains a major focus of cell cycle and cancer research. CRISPR-based models and quantitative imaging continue to reveal how cells position, assemble and constrict the division machinery.
References
- 1. Cullati SN et al.. 2019. Spatiotemporal regulation of the Dma1-mediated mitotic checkpoint coordinates mitosis with cytokinesis.. Curr Genet 65(3):663-668 PMID: 30600396
- 2. Lindon C. 2008. Control of mitotic exit and cytokinesis by the APC/C.. Biochem Soc Trans 36(Pt 3):405-10 PMID: 18481969
- 3. Holder J et al.. 2019. Getting out of mitosis: spatial and temporal control of mitotic exit and cytokinesis by PP1 and PP2A.. FEBS Lett 593(20):2908-2924 PMID: 31494926
- 4. Li JJ et al.. 2006. Mitotic kinases: the key to duplication, segregation, and cytokinesis errors, chromosomal instability, and oncogenesis.. Pharmacol Ther 111(3):974-84 PMID: 16603252
- 5. Sugioka K. 2022. Symmetry-breaking of animal cytokinesis.. Semin Cell Dev Biol 127:100-109 PMID: 34955355
- 6. Petronczki M et al.. 2008. Polo on the Rise-from Mitotic Entry to Cytokinesis with Plk1.. Dev Cell 14(5):646-59 PMID: 18477449
- 7. Sayyad WA et al.. 2022. The number of cytokinesis nodes in mitotic fission yeast scales with cell size.. Elife 11 PMID: 36093997
- 8. Kunduri G et al.. 2022. Lipid Polarization during Cytokinesis.. Cells 11(24) PMID: 36552741
