GO:1903673 mitotic cleavage furrow formation: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1903673 (mitotic cleavage furrow formation) is the biological process by which a dividing cell assembles and positions the cleavage furrow specifically during mitosis.
• The process is driven by the RhoA GTPase and its downstream effector anillin, which recruit actin and myosin to the equatorial cortex.
• Centralspindlin, a complex of MKLP1 and MgcRacGAP, is the key signaling hub that links the mitotic spindle to furrow formation.
• Rappaport's classic experiments showed that the mitotic apparatus can induce repeated furrows in sand dollar eggs, establishing the concept of cleavage signaling.
• Defects in cleavage furrow formation lead to cytokinesis failure, which can cause aneuploidy and is linked to cancer and developmental disorders.
• Research on this process uses live-cell imaging, CRISPR knockout models, and proteomics to dissect the molecular machinery.
Description
Mitotic cleavage furrow formation (GO:1903673) is the biological process that physically divides a mitotic cell into two daughter cells. It is the first visible step of cytokinesis, during which the cell cortex at the equator of the mitotic spindle invaginates to form a furrow. This process is essential for genome stability, as failure to form a proper furrow results in binucleation or aneuploidy, which are hallmarks of cancer and developmental defects. The furrow is positioned by signals from the mitotic spindle, a concept first demonstrated by Rappaport in sand dollar eggs, where a single mitotic apparatus could induce repeated furrow formation. Understanding GO:1903673 is therefore critical for researchers studying cell division, cytoskeletal dynamics, and diseases linked to cytokinesis failure.
mitotic cleavage furrow formation At A Glance
| GO ID | GO:1903673 |
|---|---|
| GO term | mitotic cleavage furrow formation |
| Ontology | biological_process |
| Synonym | cleavage furrow positioning involved in mitotic cell cycle |
| Definition | Any cleavage furrow formation that is involved in mitotic cell cycle. |
| Major function | Assembly and positioning of the cleavage furrow during mitosis, leading to cytokinesis. |
| Key regulators | RhoA, anillin, centralspindlin (MKLP1/MgcRacGAP), ECT2. |
| Cellular location | Equatorial cell cortex, actomyosin ring. |
| Related processes | Cytokinesis, actomyosin ring assembly, mitotic spindle positioning. |
What Is GO:1903673?
According to the Gene Ontology, GO:1903673 (mitotic cleavage furrow formation) is defined as any cleavage furrow formation that is involved in the mitotic cell cycle. In other words, it is the specific set of cellular events that create the cleavage furrow during mitosis, as opposed to other types of cell division. This process includes the positioning, assembly, and initial ingression of the furrow at the equatorial cortex, driven by the RhoA signaling pathway and actomyosin contractility.
Why Is mitotic cleavage furrow formation Important in Cell Biology?
Mitotic cleavage furrow formation is essential for the faithful segregation of genetic material during cell division. When this process fails, cells can become binucleated or aneuploid, which contributes to tumorigenesis and developmental disorders. Moreover, the molecular machinery of furrow formation is a target for anticancer drugs, and understanding its regulation provides insights into fundamental cell biology.
• Ensures equal distribution of chromosomes and cytoplasmic contents to daughter cells.
• Prevents aneuploidy, a hallmark of many cancers.
• Involved in tissue morphogenesis and development.
• Provides a model for studying cytoskeletal dynamics and force generation.
• Dysregulation is linked to cytokinesis failure and multinucleation in cancer cells.
• Key proteins are potential targets for chemotherapy.
• Understanding furrow positioning informs synthetic biology and regenerative medicine.
• Conserved mechanism from Dictyostelium to humans.
• Required for stem cell self-renewal and differentiation.
• Mutations in furrow components cause developmental syndromes.
What Happens During mitotic cleavage furrow formation?
Initiation and positioning of the cleavage furrow
In simple terms: The cell decides where to pinch in two, guided by the mitotic spindle.
The cleavage furrow is positioned at the equatorial cortex, equidistant from the two spindle poles. This positioning is directed by the mitotic spindle through a mechanism known as cleavage signaling, first demonstrated by Rappaport in sand dollar eggs. The central spindle, a bundle of antiparallel microtubules, recruits centralspindlin (MKLP1 and MgcRacGAP) to the midzone, which then activates the small GTPase RhoA at the equatorial cortex. This activation is the key trigger for furrow formation.
Activation of RhoA and actin polymerization
In simple terms: A molecular switch called RhoA turns on, telling the cell to build a contractile ring.
RhoA is activated by the guanine nucleotide exchange factor ECT2, which is recruited by centralspindlin. Active RhoA then activates formin proteins (e.g., mDia1) to nucleate actin filaments and Rho-kinase (ROCK) to activate myosin II. This leads to the assembly of a contractile actomyosin ring at the equatorial cortex. Anillin, an actin-binding protein, crosslinks actin and myosin and anchors the ring to the plasma membrane.
Furrow ingression and constriction
In simple terms: The ring tightens like a drawstring, pinching the cell in two.
Once assembled, the actomyosin ring contracts through the sliding of myosin II filaments along actin, generating the force that invaginates the plasma membrane. This ingression is accompanied by targeted membrane addition to accommodate the increasing surface area. The furrow deepens until it meets the midbody, leading to abscission and separation of the two daughter cells.
Coordination with mitotic exit
In simple terms: The pinching process is timed to start only after chromosomes have been separated.
Cleavage furrow formation is tightly coupled to mitotic exit. The spindle assembly checkpoint ensures that furrow formation begins only after all chromosomes are properly attached and segregated. This coordination prevents premature cytokinesis and genomic instability. Key regulators include the Aurora B kinase, which phosphorylates centralspindlin and other components to control the timing and location of furrow formation.
Key Genes Involved in GO:1903673 mitotic cleavage furrow formation
The following genes and proteins are central to mitotic cleavage furrow formation, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| RHOA | Small GTPase that activates formins and ROCK to drive actomyosin ring assembly | Key regulator; mutations linked to cancer and developmental disorders |
| ECT2 | Guanine nucleotide exchange factor that activates RhoA at the equatorial cortex | Essential for furrow initiation; target for cytokinesis research |
| KIF23 (MKLP1) | Kinesin motor subunit of centralspindlin; bundles microtubules and recruits ECT2 | Central to cleavage signaling; knockout causes furrow failure |
| RACGAP1 (MgcRacGAP) | Rho GTPase-activating protein subunit of centralspindlin; regulates RhoA activity | Required for centralspindlin function; mutations affect cytokinesis |
| ANLN (Anillin) | Actin-binding protein that crosslinks actin and myosin and anchors the ring to the membrane | Critical for furrow stability; overexpression in cancers |
| ROCK1 | Rho kinase that activates myosin II by phosphorylation | Effector of RhoA; inhibitor studies block furrow formation |
| DIAPH1 (mDia1) | Formin that nucleates actin filaments for the contractile ring | Required for actin assembly; knockout impairs cytokinesis |
| MYH9 | Non-muscle myosin II heavy chain; generates contractile force | Motor protein; mutations cause platelet disorders |
| ACTB | Beta-actin; major component of the contractile ring | Structural protein; essential for furrow ingression |
| AURKB | Aurora B kinase; regulates centralspindlin and furrow positioning | Key mitotic kinase; inhibitor targets in cancer |
| PLK1 | Polo-like kinase 1; regulates RhoA activation and furrow formation | Mitotic regulator; drug target |
| CDK1 | Cyclin-dependent kinase 1; controls mitotic entry and exit | Master regulator; inhibition blocks furrow formation |
| RhoGAP | Rho GTPase-activating proteins; fine-tune RhoA activity | Regulatory proteins; ensure proper furrow positioning |
| CIT (Citron kinase) | Serine/threonine kinase that localizes to the cleavage furrow and regulates ingression | Required for abscission; mutations cause microcephaly |
| SEPT9 | Septin filament component; stabilizes the cleavage furrow | Cytoskeletal organizer; implicated in cancer |
| ALIX | ESCRT-III accessory protein; involved in abscission | Late cytokinesis factor; knockout causes multinucleation |
| CHMP4B | ESCRT-III subunit; mediates membrane scission during abscission | Required for final separation; mutations cause cataracts |
How Is mitotic cleavage furrow formation Regulated?
Mitotic cleavage furrow formation is regulated by multiple signaling pathways. The small GTPase RhoA is the master regulator, cycling between active GTP-bound and inactive GDP-bound states. Its activation is controlled by ECT2, which is recruited by centralspindlin and phosphorylated by Aurora B and Plk1. Conversely, RhoGAPs such as MgcRacGAP and p190RhoGAP inactivate RhoA to ensure proper spatial and temporal control. Additionally, the spindle assembly checkpoint and CDK1-cyclin B activity coordinate furrow formation with chromosome segregation. Post-translational modifications, including phosphorylation and ubiquitination, fine-tune the stability and localization of furrow components.
mitotic cleavage furrow formation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ANLN | Cancer (breast, lung, liver); overexpression promotes proliferation | Knockout in cancer cell lines; overexpression in normal cells |
| ECT2 | Cancer; oncogenic when overexpressed | CRISPR knockout in HeLa cells; xenograft models |
| CIT | Microcephaly; cytokinesis failure in neural progenitors | Knockout mouse models; patient-derived iPSCs |
| MYH9 | May-Hegglin anomaly; platelet disorder | Point mutation knock-in mice; patient cells |
| RACGAP1 | Cancer; required for cytokinesis | Knockout in cancer cell lines; conditional knockout mice |
Cancer and aneuploidy
Defects in mitotic cleavage furrow formation lead to cytokinesis failure, resulting in binucleated or multinucleated cells. This genomic instability is a hallmark of many cancers, including breast, colon, and liver cancer. Overexpression of anillin (ANLN) and ECT2 has been observed in various tumors and correlates with poor prognosis. Targeting the RhoA pathway is a potential therapeutic strategy.
Developmental disorders
Mutations in genes encoding furrow components cause developmental syndromes. For example, mutations in CIT (citron kinase) cause microcephaly, and mutations in MYH9 cause May-Hegglin anomaly, a platelet disorder. These conditions highlight the importance of cleavage furrow formation in tissue development and homeostasis.
Neurodegeneration
Emerging evidence links cytokinesis defects to neurodegenerative diseases. Abnormal cleavage furrow formation can lead to aneuploid neurons, which are observed in Alzheimer's disease and other neurodegenerative conditions. However, the exact mechanisms remain under investigation.
From mitotic cleavage furrow formation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does knockout of RHOA block cleavage furrow formation? | CRISPR knockout in HeLa or MCF-7 cells |
| What is the effect of a point mutation in ECT2 on RhoA activation? | Point mutation knock-in via CRISPR in U2OS cells |
| Can we visualize centralspindlin dynamics in live cells? | Knock-in of fluorescent tags (e.g., GFP) on KIF23 |
| Does overexpression of ANLN cause multinucleation? | Overexpression of ANLN in normal fibroblasts |
| What is the role of CIT in neural development? | Conditional knockout in mouse neural progenitors |
| Can CRISPR library screening identify new furrow regulators? | Genome-wide knockout library in haploid HAP1 cells |
How to Study the mitotic cleavage furrow formation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Live-cell imaging | Dynamics of furrow formation and ingression | Visualizing actin and myosin recruitment |
| CRISPR knockout | Gene requirement for furrow formation | Testing essential genes in cell lines |
| CRISPR point mutation | Effect of specific mutations on protein function | Studying disease-associated variants |
| Proteomics | Protein interactions and modifications | Mapping the furrow interactome |
| RNA-seq | Transcriptional changes during mitosis | Identifying upregulated furrow genes |
| High-content screening | Phenotypic defects (multinucleation) | Genome-wide screens for novel regulators |
| FRET biosensors | RhoA activity at the equatorial cortex | Spatiotemporal analysis of signaling |
| Electron microscopy | Ultrastructure of the cleavage furrow | Detailed morphology of the contractile ring |
Live-cell imaging
Live-cell imaging with fluorescently tagged proteins (e.g., GFP-actin, mCherry-myosin) allows real-time visualization of cleavage furrow formation and ingression. This method is essential for studying the dynamics and timing of furrow assembly.
CRISPR knockout and point mutation
CRISPR-Cas9 technology enables the generation of knockout cell lines to test the requirement of specific genes in furrow formation. Point mutations can be introduced to study the effects of specific amino acid changes on protein function.
Proteomics and interactomics
Mass spectrometry-based proteomics can identify protein-protein interactions and post-translational modifications of furrow components. This approach helps map the signaling network of cleavage furrow formation.
High-content screening
High-content screening with siRNA or CRISPR libraries can identify novel regulators of cytokinesis. Automated imaging and analysis quantify multinucleation and furrow defects.
How CRISPR Can Be Used to Study GO:1903673 mitotic cleavage furrow formation
Knockout
CRISPR knockout of genes such as RHOA, ECT2, or KIF23 results in failure of cleavage furrow formation, leading to binucleated cells. These models are used to confirm the essential role of these genes in cytokinesis.
Point Mutation
Point mutations can be introduced to mimic disease-associated variants or to dissect domain functions. For example, a point mutation in the RhoA-binding domain of anillin can disrupt its localization to the furrow.
Knock-in
Knock-in of fluorescent tags (e.g., GFP, mCherry) allows live-cell imaging of endogenous proteins. This is particularly useful for tracking centralspindlin dynamics during furrow formation.
Overexpression
Overexpression of furrow components such as ANLN or ECT2 can induce ectopic furrow formation or multinucleation, providing insights into their oncogenic potential.
How EDITGENE Supports mitotic cleavage furrow formation Research
Researchers studying mitotic cleavage furrow formation-related genes often need to determine whether a candidate gene is causally involved in the process or merely correlated. This requires precise genetic manipulation, which can be achieved through CRISPR-based knockout, point mutation, knock-in, or overexpression models. EDITGENE provides these services to accelerate your research.
Contact EDITGENE today to design your custom CRISPR model for mitotic cleavage furrow formation research.
Frequently Asked Questions About mitotic cleavage furrow formation
What is mitotic cleavage furrow formation?
Mitotic cleavage furrow formation (GO:1903673) is the process by which a cell assembles and positions the cleavage furrow during mitosis, leading to cytokinesis.
What genes are involved in mitotic cleavage furrow formation?
Key genes include RHOA, ECT2, KIF23, RACGAP1, ANLN, and CIT, among others.
How is the cleavage furrow positioned?
The mitotic spindle signals to the equatorial cortex via centralspindlin and RhoA to position the furrow.
What happens if cleavage furrow formation fails?
Failure leads to binucleation or multinucleation, which can cause aneuploidy and is linked to cancer.
What is the role of RhoA in cleavage furrow formation?
RhoA is a small GTPase that activates formins and ROCK to assemble the actomyosin contractile ring.
How can I study mitotic cleavage furrow formation?
Common methods include live-cell imaging, CRISPR knockout, proteomics, and high-content screening.
What is centralspindlin?
Centralspindlin is a complex of MKLP1 and MgcRacGAP that links the central spindle to RhoA activation at the furrow.
Is mitotic cleavage furrow formation conserved?
Yes, the core machinery is conserved from Dictyostelium to humans.
What diseases are associated with defects in cleavage furrow formation?
Cancer, microcephaly, and platelet disorders such as May-Hegglin anomaly.
Can CRISPR be used to study cleavage furrow formation?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to dissect gene function.
Conclusion
Mitotic cleavage furrow formation (GO:1903673) is a fundamental biological process that ensures accurate cell division. Its molecular machinery, centered on RhoA and centralspindlin, is highly conserved and tightly regulated. Defects in this process contribute to cancer and developmental disorders, making it a critical area of research. Advances in CRISPR technology and imaging are accelerating our understanding of this process, and EDITGENE provides the tools to support these studies.
References
- 1. Satterwhite LL et al.. 1992. Cytokinesis.. Curr Opin Cell Biol 4(1):43-52 PMID: 1313686
- 2. Glotzer M. 2004. Cleavage furrow positioning.. J Cell Biol 164(3):347-51 PMID: 14757750
- 3. Scholey JM. 2025. Mitotic spindle membranes.. Mol Biol Cell 36(4):re1 PMID: 40067152
- 4. Earnshaw WC et al.. 1994. Mitosis.. Bioessays 16(9):639-43 PMID: 7980489
- 5. Okada A et al.. 2023. Cleavage furrow positioning in dividing Dictyostelium cells.. Cytoskeleton (Hoboken) 80(11-12):448-460 PMID: 37650534
- 6. D'Avino PP et al.. 2005. Cleavage furrow formation and ingression during animal cytokinesis: a microtubule legacy.. J Cell Sci 118(Pt 8):1549-58 PMID: 15811947
- 7. Rappaport R. 1985. Repeated furrow formation from a single mitotic apparatus in cylindrical sand dollar eggs.. J Exp Zool 234(1):167-71 PMID: 3989496
- 8. Mishima M. 2016. Centralspindlin in Rappaport's cleavage signaling.. Semin Cell Dev Biol 53:45-56 PMID: 26964770
