GO:1905345 protein localization to cleavage furrow: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1905345 (protein localization to cleavage furrow) describes the directed transport or retention of proteins at the cleavage furrow, the equatorial actomyosin-rich zone that divides a cell during cytokinesis.
• The process depends on cytoskeletal motors, membrane trafficking, and lipid delivery; for example, Kif12 is required for INCENP localization at the furrow, and the FIP3-Rab11 complex targets recycling endosomes to the furrow.
• Nonmuscle myosin II isoform activity determines myosin localization at the cleavage furrow in megakaryocytes, linking this GO term to polyploidization.
• In some organisms, furrow formation and protein recruitment can occur without F-actin, as shown in Chlamydomonas, indicating conserved but mechanistically diverse pathways.
• Lipid delivery through the endocytic pathway, such as ceramide phosphoethanolamine transport, is essential for male meiotic cytokinesis and for proper furrow protein/lipid composition.
• Dysregulation of cleavage furrow protein localization contributes to cytokinesis failure, aneuploidy, and diseases such as cancer and blood disorders.
Description
Cytokinesis is the final step of cell division, during which a cleavage furrow forms and ingresses to separate the daughter cells. The spatial and temporal control of this process requires the precise localization of numerous proteins to the cleavage furrow. GO:1905345, protein localization to cleavage furrow, is the biological process that encompasses the transport or maintenance of proteins at this specialized equatorial region. Understanding this process is fundamental to cell biology because it ensures the fidelity of chromosome segregation and cell division. Defects in protein localization to the cleavage furrow can lead to cytokinesis failure, resulting in binucleation or aneuploidy, which are hallmarks of cancer and other proliferative disorders. Moreover, the machinery that localizes proteins to the furrow is conserved across eukaryotes, from Dictyostelium to humans, making it a rich area for comparative and mechanistic studies. Researchers study this term using advanced imaging, proteomics, and genetic perturbation, and it is increasingly recognized as a hub for signaling and membrane trafficking events.
protein localization to cleavage furrow At A Glance
| GO ID | GO:1905345 |
|---|---|
| GO term | protein localization to cleavage furrow |
| Ontology | biological_process |
| Synonym | protein localisation in cleavage furrow; protein localisation to cleavage furrow; protein localization in cleavage furrow |
| Major function | Transport or maintenance of proteins at the cleavage furrow during cytokinesis |
| Related cellular component | cleavage furrow |
| Related biological process | cytokinesis; actomyosin contractile ring assembly |
| Key molecules | INCENP, Kif12, Rab11, FIP3, nonmuscle myosin II, ceramide phosphoethanolamine |
What Is GO:1905345?
According to the Gene Ontology, GO:1905345 (protein localization to cleavage furrow) is defined as a process in which a protein is transported to, or maintained in, a location within a cleavage furrow. This includes both the active delivery of proteins to the furrow and the mechanisms that retain them there. The term is a biological process and is synonymous with protein localisation in cleavage furrow, protein localisation to cleavage furrow, and protein localization in cleavage furrow.
Why Is protein localization to cleavage furrow Important in Cell Biology?
Protein localization to the cleavage furrow is essential for cytokinesis, the process that physically separates daughter cells after mitosis. Without the correct spatiotemporal deposition of proteins such as INCENP, myosin II, and Rab11 effectors, the furrow fails to ingress properly, leading to cytokinesis failure, aneuploidy, and cell death or transformation. This process is also critical for specialized cell divisions, including megakaryocyte polyploidization and male meiosis, where defects can cause thrombocytopenia or infertility. Thus, understanding GO:1905345 provides mechanistic insight into fundamental cell division and offers potential therapeutic targets for diseases characterized by aberrant proliferation.
• Ensures faithful cytokinesis and prevents aneuploidy, a driver of cancer and developmental disorders.
• Regulates megakaryocyte polyploidization, which is necessary for platelet production; defects can lead to thrombocytopenia.
• Required for male meiotic cytokinesis, with lipid delivery defects causing infertility.
• Coordinates membrane trafficking with actomyosin ring contraction, linking endocytic pathways to cell division.
• Involved in diverse organisms, from Dictyostelium to humans, highlighting evolutionary conservation.
• Provides targets for anti-cancer strategies aimed at cytokinesis failure.
• Serves as a model for studying spatial regulation of protein localization and signaling.
• Implicated in Chlamydomonas cell division, where F-actin-independent furrowing occurs, expanding mechanistic understanding.
• Links to lipid metabolism and endocytic recycling, integrating membrane dynamics with cytoskeletal function.
• Offers biomarkers for cytokinesis-related pathologies when localization is perturbed.
What Happens During protein localization to cleavage furrow?
Initiation and specification of the cleavage furrow
In simple terms: The cell decides where to pinch in two.
The cleavage furrow is specified at the equatorial region of the cell, often through signals from the mitotic spindle. This involves the activation of RhoA and the assembly of an actomyosin contractile ring. Proteins such as INCENP, a chromosomal passenger complex component, are among the first to localize to the furrow, and this localization depends on the kinesin Kif12 and interactions with the actin cytoskeleton. In Dictyostelium, a mitotic kinesin-like protein is required for normal karyokinesis and for myosin localization to the furrow, indicating that motor proteins play an early role in furrow specification.
Active transport of proteins to the furrow
In simple terms: Molecular motors carry proteins to the pinch site.
Once the furrow is specified, proteins are actively transported along cytoskeletal tracks. The FIP3-Rab11 protein complex regulates the targeting of recycling endosomes to the cleavage furrow during late cytokinesis, delivering proteins and lipids to the equatorial membrane. Similarly, Kif12 is required for the localization of INCENP at the cleavage furrow, and this involves interactions of the INCENP N-terminus with the actin cytoskeleton. These transport mechanisms ensure that the furrow receives the necessary components for ingression.
Retention and maintenance at the furrow
In simple terms: Proteins are held in place at the pinch site.
Localization is not only about delivery but also about keeping proteins at the furrow. Nonmuscle myosin II isoforms exhibit differential activity that determines their localization at the cleavage furrow of megakaryocytes, and this retention is critical for proper furrow ingression. Theoretical work suggests that static cytokinetic furrows can arise from traveling excitable waves, which may help maintain protein gradients at the furrow. Thus, retention mechanisms counteract diffusion and ensure a stable furrow proteome.
Lipid and membrane contribution to protein localization
In simple terms: Fats help bring proteins to the pinch.
Membrane lipids are not passive; they actively participate in protein localization. Delivery of ceramide phosphoethanolamine lipids to the cleavage furrow through the endocytic pathway is essential for male meiotic cytokinesis. This lipid transport likely creates a specialized membrane domain that recruits or stabilizes furrow proteins. The FIP3-Rab11 complex also delivers membrane to the furrow, linking lipid and protein trafficking.
F-actin-independent mechanisms
In simple terms: Some cells can pinch without the usual actin cables.
While actomyosin is central in many systems, cleavage-furrow formation can occur without F-actin in Chlamydomonas, indicating alternative mechanisms for furrow assembly and protein localization. This suggests that the core machinery for protein localization to the furrow may be more diverse than previously thought, and that other cytoskeletal or membrane-based forces can drive furrowing.
Key Genes Involved in GO:1905345 protein localization to cleavage furrow
The following genes and proteins are experimentally implicated in protein localization to the cleavage furrow, based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| INCENP | Chromosomal passenger complex protein; localizes to cleavage furrow | Its furrow localization depends on Kif12 and actin; key marker for cytokinesis |
| Kif12 | Kinesin motor protein; required for INCENP furrow localization | Links motor activity to chromosomal passenger complex targeting |
| Rab11 | Small GTPase; regulates recycling endosome targeting to furrow | Part of FIP3-Rab11 complex; controls membrane delivery during late cytokinesis |
| FIP3 | Rab11 effector; mediates endosome targeting to furrow | Essential for furrow membrane trafficking |
| Myosin II (nonmuscle) | Contractile ring component; localizes to furrow | Isoform-specific activity determines furrow localization in megakaryocytes |
| RhoA | GTPase; regulates actomyosin ring assembly | Upstream regulator of furrow formation (implied by) |
| Anillin | Actin-binding protein; links cytoskeleton to membrane at furrow | Not directly cited but commonly studied in furrow context |
| Septins | Filament-forming proteins; localize to furrow | Not directly cited but implicated in cytokinesis |
| CPE (ceramide phosphoethanolamine) | Lipid; delivered to furrow via endocytic pathway | Essential for male meiotic cytokinesis |
| Kinesin-like protein (Dictyostelium) | Mitotic kinesin; required for myosin localization to furrow | Shows motor-dependent furrow protein targeting |
| Actin | Cytoskeletal filament; interacts with INCENP N-terminus | Required for INCENP furrow localization |
| EHD1 | Membrane trafficking protein; involved in recycling endosome delivery | Not directly cited but related to Rab11 pathway |
| MgcRacGAP | Rho GTPase-activating protein; central spindle and furrow | Not directly cited but key regulator |
| Aurora B | Kinase; part of chromosomal passenger complex | Not directly cited but co-localizes with INCENP |
| Survivin | Chromosomal passenger complex component | Not directly cited but partners with INCENP |
| Borealin | Chromosomal passenger complex component | Not directly cited but partners with INCENP |
| PRC1 | Microtubule bundling protein; central spindle | Not directly cited but influences furrow positioning |
| Citron kinase | Rho effector; contractile ring | Not directly cited but involved in furrow ingression |
How Is protein localization to cleavage furrow Regulated?
The process of protein localization to the cleavage furrow is regulated by multiple mechanisms. The small GTPase Rab11, in complex with FIP3, controls the timing and targeting of recycling endosomes to the furrow during late cytokinesis. Nonmuscle myosin II isoform-specific activity determines the localization of myosin itself at the furrow, suggesting that motor activity and post-translational modifications regulate retention. In megakaryocytes, polyploidization requires precise regulation of myosin localization, which is linked to cell cycle and differentiation signals. Additionally, theoretical models propose that traveling excitable waves of signaling molecules can create static furrows, implying that reaction-diffusion systems regulate protein distribution. Lipid delivery, such as ceramide phosphoethanolamine transport, is also a regulatory input for furrow function in meiosis.
protein localization to cleavage furrow and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| INCENP | Cancer, aneuploidy | KO or point mutation in cancer cell lines; imaging of furrow |
| Kif12 | Cytokinesis defects, potential developmental disorders | KO in HeLa or RPE1 cells; rescue with tagged Kif12 |
| Rab11 | Cancer, membrane trafficking disorders | Knock-in of GFP-Rab11; live imaging |
| Myosin II | Thrombocytopenia, bleeding disorders | Megakaryocyte KO; polyploidization assays |
| Ceramide phosphoethanolamine synthase | Male infertility | Testis-specific KO in mice; meiotic spread assays |
Cancer and aneuploidy
Failure of protein localization to the cleavage furrow leads to cytokinesis defects, resulting in binucleation and aneuploidy, which are hallmarks of cancer. For example, mislocalization of INCENP or myosin II can cause furrow regression and genomic instability. Targeting these localization pathways is a potential anti-cancer strategy.
Hematological disorders
In megakaryocytes, proper myosin II localization at the cleavage furrow is required for polyploidization, a process essential for platelet production. Defects in this localization can lead to thrombocytopenia or abnormal platelet function.
Male infertility
Male meiotic cytokinesis requires the delivery of specific lipids, such as ceramide phosphoethanolamine, to the cleavage furrow. Disruption of this lipid transport leads to cytokinesis failure and infertility in model organisms.
From protein localization to cleavage furrow-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X localize to the cleavage furrow? | Tagged knock-in (e.g., GFP) in HeLa or RPE1 cells; live imaging |
| Is gene X required for furrow protein localization? | CRISPR knockout followed by immunofluorescence of furrow markers |
| Does a disease-associated mutation affect furrow localization? | Point mutation knock-in via CRISPR; compare to wild-type |
| Can overexpression of gene X rescue localization defects? | Overexpression construct in KO background; rescue imaging |
| What proteins interact with gene X at the furrow? | Knock-in of proximity labeling tags (BioID, APEX); proteomics |
| Is gene X involved in lipid-dependent furrow targeting? | KO in meiotic cells; lipid supplementation assays |
How to Study the protein localization to cleavage furrow Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Live-cell fluorescence imaging | Dynamic localization of tagged proteins | Tracking INCENP or Rab11 to furrow |
| Immunofluorescence | Steady-state localization of endogenous proteins | Quantifying myosin II at furrow in KO cells |
| Proximity labeling (BioID/APEX) | Protein interactome at furrow | Identifying novel furrow components |
| CRISPR knockout | Requirement of gene for furrow localization | Testing Kif12 role in INCENP targeting |
| RNAi/knockdown | Acute depletion effects | Validating motor protein function |
| Lipidomics | Lipid composition at furrow | Ceramide phosphoethanolamine delivery |
| Theoretical modeling | Reaction-diffusion dynamics | Explaining static furrow from waves |
| Correlative light-electron microscopy | Ultrastructure of furrow | Visualizing membrane trafficking |
Live-cell imaging
Fluorescently tagged proteins (e.g., GFP-INCENP, GFP-Rab11) can be imaged in dividing cells to track their localization to the cleavage furrow in real time. This method reveals dynamics of delivery and retention.
Immunofluorescence and fixed-cell microscopy
Fixed cells stained with antibodies against furrow proteins (e.g., myosin II, INCENP) allow quantification of localization defects in knockout or mutant backgrounds.
Proteomics and proximity labeling
BioID or APEX2 fused to furrow proteins can identify nearby proteins and reveal the furrow proteome. This helps uncover novel components involved in localization.
Genetic perturbation and rescue
CRISPR knockout of candidate genes followed by rescue with wild-type or mutant constructs tests necessity and sufficiency for furrow localization.
How CRISPR Can Be Used to Study GO:1905345 protein localization to cleavage furrow
Knockout
CRISPR knockout of genes such as Kif12 or Rab11 can abolish protein localization to the cleavage furrow, revealing essential roles. For example, Kif12 knockout prevents INCENP furrow localization, and Rab11 knockout disrupts endosome targeting.
Point Mutation
Introducing disease-associated or phospho-mimetic point mutations into genes like myosin II can test how specific residues affect furrow localization. This is useful for dissecting regulatory phosphorylation.
Knock-in
Knock-in of fluorescent tags (e.g., GFP) or proximity labeling enzymes (BioID) at endogenous loci allows real-time tracking and interactome mapping of furrow proteins without overexpression artifacts.
Overexpression
Overexpression of wild-type or mutant proteins can test sufficiency for furrow localization and rescue of knockout phenotypes. For instance, overexpressing INCENP mutants can map domains required for furrow targeting.
How EDITGENE Supports protein localization to cleavage furrow Research
Researchers studying protein localization to cleavage furrow-related genes often need to determine whether a candidate gene is causally involved in the process or is merely a bystander. This requires precise genetic models that can knock out, mutate, tag, or overexpress the gene of interest in relevant cell types. EDITGENE provides a comprehensive suite of CRISPR services to accelerate such studies.
Contact EDITGENE today to design your custom CRISPR model for protein localization to cleavage furrow research.
Frequently Asked Questions About protein localization to cleavage furrow
What is protein localization to cleavage furrow?
It is the biological process (GO:1905345) by which proteins are transported to or maintained at the cleavage furrow during cytokinesis.
What genes are involved in protein localization to cleavage furrow?
Key genes include INCENP, Kif12, Rab11, FIP3, and nonmuscle myosin II, among others.
Why is protein localization to the cleavage furrow important?
It ensures proper cytokinesis; defects cause aneuploidy, cancer, and blood disorders.
How is protein localization to the cleavage furrow regulated?
It is regulated by Rab11-FIP3 endosome targeting, myosin II activity, and lipid delivery.
What diseases are associated with defects in cleavage furrow protein localization?
Cancer, thrombocytopenia, and male infertility are linked to defects in this process.
What methods are used to study protein localization to the cleavage furrow?
Live imaging, immunofluorescence, proteomics, and CRISPR knockout are common methods.
Can protein localization to the cleavage furrow occur without F-actin?
Yes, in Chlamydomonas, furrow formation can occur without F-actin, indicating alternative mechanisms.
What is the role of Kif12 in cleavage furrow localization?
Kif12 is required for INCENP localization at the furrow, linking motor activity to chromosomal passenger complex targeting.
How does Rab11 regulate cleavage furrow protein localization?
Rab11, with FIP3, targets recycling endosomes to the furrow, delivering proteins and lipids during late cytokinesis.
What is the role of myosin II in cleavage furrow localization?
Nonmuscle myosin II isoform activity determines its own localization at the furrow, essential for ingression.
Conclusion
Protein localization to the cleavage furrow (GO:1905345) is a fundamental biological process that ensures the precise delivery and retention of proteins at the equatorial region during cytokinesis. Research has identified key molecular players such as INCENP, Kif12, Rab11, FIP3, and myosin II, and has revealed diverse mechanisms including motor-dependent transport, membrane trafficking, and lipid delivery. Defects in this process are linked to cancer, hematological disorders, and infertility, making it a compelling area for therapeutic intervention. Continued investigation using advanced CRISPR models and imaging will further illuminate the regulatory networks and translational potential of this process.
References
- 1. Mazzi S et al.. 2018. Megakaryocyte and polyploidization.. Exp Hematol 57:1-13 PMID: 29111429
- 2. Chen Q et al.. 2007. The localization of inner centromeric protein (INCENP) at the cleavage furrow is dependent on Kif12 and involves interactions of the N terminus of INCENP with the actin cytoskeleton.. Mol Biol Cell 18(9):3366-74 PMID: 17567958
- 3. Wilson GM et al.. 2005. The FIP3-Rab11 protein complex regulates recycling endosome targeting to the cleavage furrow during late cytokinesis.. Mol Biol Cell 16(2):849-60 PMID: 15601896
- 4. Onishi M et al.. 2020. Cleavage-furrow formation without F-actin in Chlamydomonas.. Proc Natl Acad Sci U S A 117(31):18511-18520 PMID: 32690698
- 5. Kunduri G et al.. 2022. Delivery of ceramide phosphoethanolamine lipids to the cleavage furrow through the endocytic pathway is essential for male meiotic cytokinesis.. PLoS Biol 20(9):e3001599 PMID: 36170207
- 6. Goryachev AB et al.. 2016. How to make a static cytokinetic furrow out of traveling excitable waves.. Small GTPases 7(2):65-70 PMID: 27070950
- 7. Roy A et al.. 2016. Activity of nonmuscle myosin II isoforms determines localization at the cleavage furrow of megakaryocytes.. Blood 128(26):3137-3145 PMID: 27737892
- 8. Lakshmikanth GS et al.. 2004. A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium.. Proc Natl Acad Sci U S A 101(47):16519-24 PMID: 15546981
