GO:0051010 microtubule plus-end binding: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0051010 microtubule plus-end binding describes the biological process in which proteins selectively recognize and associate with the growing plus ends of microtubules.
• Plus-end tracking proteins (+TIPs) such as EB1, EB3, CLIP-170, CLASPs, and TACC3 form dynamic comet-like accumulations at microtubule plus ends.
• Phase separation of EB1 and its partners has emerged as a key mechanism guiding plus-end dynamics and mitotic spindle organization.
• Dysregulation of plus-end binding contributes to cancer, neurodevelopmental disorders, and mitotic defects.
• CRISPR knockout, point-mutation, knock-in, and overexpression models enable causal dissection of plus-end binding genes.
• Advanced imaging, proteomics, and CRISPR library screening are standard methods for studying microtubule plus-end binding.
Description
Microtubules are dynamic cytoskeletal polymers whose plus ends explore cellular space and mediate chromosome segregation, intracellular transport, and cell polarity. The biological process GO:0051010, microtubule plus-end binding, refers to the selective interaction of proteins with the growing plus ends of microtubules, often producing comet-like accumulations that track with polymerization. These plus-end tracking proteins, or +TIPs, are not merely passive passengers; they regulate microtubule dynamics, link microtubules to cellular structures, and coordinate force generation during mitosis and migration. Understanding this process is essential because it sits at the intersection of cytoskeletal regulation, cell division, and neuronal development. Defects in plus-end binding factors are increasingly linked to human disease, including cancer and neurodevelopmental conditions. This article integrates authoritative GO annotation with verified PubMed literature to provide a research-grade overview of GO:0051010, its molecular players, regulatory logic, and the CRISPR-based models used to study it.
microtubule plus-end binding At A Glance
| GO ID | GO:0051010 |
|---|---|
| GO term | microtubule plus-end binding |
| Ontology | biological_process |
| Synonym | None listed in QuickGO |
| Major function | Selective association of proteins with growing microtubule plus ends to regulate dynamics and interactions |
| Key proteins | EB1, EB3, CLIP-170, CLASPs, TACC3, septin 9, and other +TIPs |
| Cellular context | Mitotic spindle, interphase microtubule network, neuronal growth cones |
| Related disease areas | Cancer, neurodevelopmental disorders, mitotic defects |
What Is GO:0051010?
Microtubule plus-end binding (GO:0051010) is the biological process in which a protein or protein complex selectively binds to the plus end of a microtubule, the faster-growing end that dynamically switches between growth and shrinkage. This binding is often transient and coupled to microtubule polymerization, enabling proteins to track the plus end as a comet-like structure. The process is distinct from general microtubule binding because it requires recognition of the plus-end conformation or the nucleotide state of tubulin, and it underlies the recruitment of diverse +TIP networks that control microtubule dynamics and interactions.
Why Is microtubule plus-end binding Important in Cell Biology?
Microtubule plus-end binding is fundamental to how cells organize their microtubule cytoskeleton in space and time. By concentrating regulatory proteins at growing plus ends, cells can rapidly respond to signals, position the mitotic spindle, and direct cargo transport. Disruption of plus-end tracking leads to spindle misorientation, chromosome missegregation, and defective neuronal connectivity, underscoring its importance in both basic cell biology and disease mechanisms.
• Controls microtubule dynamic instability and polymerization rates.
• Essential for mitotic spindle assembly and chromosome segregation.
• Guides neuronal development, axon guidance, and growth cone motility.
• Regulates intracellular cargo transport in interphase cells.
• Links microtubules to actin and septin filaments for cellular morphogenesis.
• Phase separation of EB1 organizes plus-end compartments.
• Dysregulation is implicated in cancer cell proliferation and metastasis.
• Mutations in plus-end proteins are linked to neurodevelopmental disorders.
• Serves as a target for anti-mitotic chemotherapeutics.
• Provides a model system for studying dynamic protein-protein interactions.
What Happens During microtubule plus-end binding?
Recognition of the growing plus end
In simple terms: Proteins find and grab the fast-growing end of a microtubule.
Plus-end binding begins when +TIPs recognize structural features of the microtubule plus end, such as the GTP-bound tubulin conformation or the open sheet at the growing tip. EB1 family proteins are among the first to bind, forming comet-like accumulations that track with polymerization. This recognition is highly dynamic and depends on the nucleotide state and curvature of the microtubule lattice.
Assembly of +TIP networks
In simple terms: Multiple proteins gather together at the plus end to form a team.
Once EB1 is bound, it recruits other +TIPs including CLIP-170, CLASPs, and TACC3 through specific binding motifs. These interactions build a dynamic network that can modulate microtubule stability and link the plus end to cellular structures. The composition of this network changes with cell cycle stage and signaling cues.
Phase separation and compartmentalization
In simple terms: Proteins can condense into droplets at the plus end to organize the area.
Recent evidence shows that EB1 and its partners undergo liquid-liquid phase separation, forming biomolecular condensates at microtubule plus ends. This phase separation guides plus-end dynamics and is regulated during mitosis to ensure proper spindle function. Such condensates concentrate regulatory factors and exclude others, creating a specialized plus-end compartment.
Regulation of microtubule dynamics
In simple terms: The plus-end team controls how fast the microtubule grows or shrinks.
Plus-end binding proteins directly influence microtubule dynamic instability by altering catastrophe and rescue frequencies. For example, TACC3 regulates plus-end dynamics and cargo transport in interphase cells, while septin 9 modulates plus-end dynamics through its interaction with microtubules. These regulatory effects are critical for spindle positioning and cell migration.
Coupling to cellular functions
In simple terms: The plus-end team connects microtubules to other cell parts to do jobs.
Plus-end binding is coupled to diverse cellular functions including chromosome capture, nuclear positioning, and neuronal growth cone guidance. In neurons, +TIPs such as EB1 and CLIP-170 regulate microtubule remodeling during axon outgrowth. In mitotic cells, plus-end tracking ensures proper kinetochore attachment and spindle assembly.
Key Genes Involved in GO:0051010 microtubule plus-end binding
The following genes encode proteins that directly bind microtubule plus ends or are core components of plus-end tracking networks, as supported by the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| MAPRE1 (EB1) | Core plus-end tracking protein; recruits other +TIPs | Phase separation, mitotic spindle, cancer |
| MAPRE2 (EB2) | Plus-end tracking; regulates microtubule dynamics | Neuronal development, cell migration |
| MAPRE3 (EB3) | Plus-end tracking; neuronal microtubule regulation | Axon guidance, growth cone |
| CLIP1 (CLIP-170) | Links plus ends to organelles and kinetochores | Mitosis, cargo transport |
| CLASP1 | Regulates microtubule stability at plus ends | Spindle positioning, neuronal polarity |
| CLASP2 | Regulates microtubule stability at plus ends | Cell migration, adhesion |
| TACC3 | Regulates plus-end dynamics and cargo transport | Interphase transport, cancer |
| SEPT9 | Modulates plus-end dynamics | Cytokinesis, cytoskeletal crosstalk |
| DCTN1 (p150glued) | Dynein adaptor at plus ends | Transport, neurodegeneration |
| KIF2A | Kinesin-13 depolymerase at plus ends | Mitotic spindle, neuronal development |
| KIF2C (MCAK) | Kinesin-13 depolymerase at plus ends | Chromosome segregation |
| CENP-E | Kinesin at kinetochores tracking plus ends | Mitosis, cancer |
| APC | Tumor suppressor tracking plus ends | Colorectal cancer, spindle orientation |
| EB1-APC complex | Regulates microtubule stability | Cancer cell migration |
| CLIP-170-EB1 | Cooperative plus-end tracking | Dynamic instability |
| TACC3-EB1 | Regulates interphase transport | Cargo trafficking |
| SEPT9-EB1 | Crosstalk with septin filaments | Cytokinesis |
How Is microtubule plus-end binding Regulated?
Microtubule plus-end binding is regulated at multiple levels. Post-translational modifications of tubulin, such as detyrosination and acetylation, can influence +TIP recruitment. Phosphorylation of EB1 and other +TIPs by mitotic kinases modulates their binding affinity and phase separation behavior during mitosis. Additionally, the availability of binding partners and the local concentration of tubulin dimers affect the size and lifetime of plus-end comets. Septin 9 has been shown to regulate plus-end dynamics, indicating crosstalk with the septin cytoskeleton. These regulatory layers ensure that plus-end tracking is responsive to cell cycle and signaling cues.
microtubule plus-end binding and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| MAPRE1 (EB1) | Cancer, chromosomal instability | Knockout and overexpression in cancer cell lines |
| TACC3 | Cancer, mitotic defects | Point mutation and knockout in HeLa cells |
| APC | Colorectal cancer | Knockout in intestinal organoids |
| CLASP2 | Neurodevelopmental disorders | Knockout in neuronal cultures |
| SEPT9 | Leukemia, cytokinesis defects | Knockdown and knockout in hematopoietic cells |
Cancer and mitotic dysregulation
Altered expression or mutation of plus-end binding proteins such as EB1, TACC3, and APC is observed in multiple cancers. EB1 overexpression correlates with poor prognosis in several tumor types, and its role in phase separation may contribute to mitotic spindle defects and chromosomal instability. TACC3 is implicated in cancer cell proliferation and is a potential therapeutic target. APC, a classic tumor suppressor, tracks plus ends to regulate spindle orientation, and its loss contributes to colorectal cancer.
Neurodevelopmental and neurodegenerative disorders
Plus-end tracking proteins are critical for neuronal microtubule remodeling. Mutations or dysregulation of EB3, CLASP2, and other +TIPs have been linked to neurodevelopmental disorders characterized by axon guidance defects and impaired neuronal migration. Disruption of plus-end binding can also contribute to neurodegeneration through defective cargo transport.
Mitotic and cytoskeletal diseases
Defects in plus-end binding can lead to cytokinesis failure and spindle misorientation, which are associated with developmental disorders and cancer. Septin 9, which regulates plus-end dynamics, is involved in cytokinesis and its dysfunction is linked to leukemia and other malignancies.
From microtubule plus-end binding-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does EB1 phase separation drive plus-end tracking? | Knock-in of phase-separation-deficient EB1 mutants |
| What is the role of TACC3 in interphase transport? | CRISPR knockout of TACC3 in HeLa cells |
| How does septin 9 regulate plus-end dynamics? | Point mutation of SEPT9 in fibroblasts |
| Is CLASP2 required for neuronal migration? | Conditional knockout in mouse neurons |
| Can EB1 overexpression induce chromosomal instability? | Overexpression in cancer cell lines |
| What is the function of APC at plus ends? | Knock-in of tagged APC in colorectal cells |
How to Study the microtubule plus-end binding Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Live-cell fluorescence microscopy | Plus-end tracking dynamics and comet formation | EB1-GFP imaging in mitotic cells |
| CRISPR knockout screening | Genes required for plus-end binding | Cancer cell line screens |
| Affinity proteomics | Protein interactions at plus ends | EB1 interactome |
| In vitro reconstitution | Binding kinetics and phase separation | Purified EB1 and tubulin |
| RNA-seq | Transcriptional changes upon +TIP perturbation | Knockout models |
| Proximity labeling | Local interactome at plus ends | APEX2-tagged EB1 |
| High-resolution imaging | Nanoscale organization of plus-end complexes | STORM/PALM |
| Mitotic spindle assays | Spindle assembly and chromosome segregation | TACC3 knockout |
Live-cell imaging of plus-end tracking
Fluorescence microscopy of GFP-tagged +TIPs such as EB1 allows real-time visualization of comet-like plus-end accumulations. This method measures microtubule growth rates, catastrophe frequencies, and the dynamics of phase-separated condensates.
Proteomics of plus-end complexes
Affinity purification coupled with mass spectrometry can identify the composition of plus-end tracking networks. This approach has revealed interactions between EB1, CLIP-170, and TACC3, and can be combined with CRISPR knockout to validate dependencies.
CRISPR library screening
Genome-wide CRISPR knockout screens can identify genes required for plus-end binding and mitotic fidelity. Such screens have uncovered novel regulators of microtubule dynamics and are applicable to cancer and neurodevelopmental models.
In vitro reconstitution and phase separation assays
Purified proteins can be reconstituted with microtubules in vitro to study binding kinetics and phase separation. These assays provide mechanistic insight into how EB1 and partners form condensates at plus ends.
How CRISPR Can Be Used to Study GO:0051010 microtubule plus-end binding
Knockout
CRISPR knockout of plus-end binding genes such as MAPRE1, TACC3, or CLASP2 enables loss-of-function studies to determine their role in microtubule dynamics, mitosis, and neuronal development. Knockout cell lines can be validated by western blot and live-cell imaging.
Point Mutation
Introducing point mutations that abolish specific interactions or phase separation (e.g., in EB1) allows precise dissection of molecular mechanisms without complete loss of protein. Such models are valuable for studying phosphorylation sites and binding interfaces.
Knock-in
Knock-in of fluorescent or affinity tags (e.g., GFP, HaloTag, APEX2) at endogenous loci enables real-time tracking and proximity proteomics of plus-end proteins under native regulation. This approach preserves endogenous expression levels and splicing.
Overexpression
Overexpression of plus-end proteins such as EB1 or TACC3 can model cancer-associated gain-of-function phenotypes, including chromosomal instability and altered cell migration. Inducible systems allow controlled expression to avoid toxicity.
How EDITGENE Supports microtubule plus-end binding Research
Researchers studying microtubule plus-end binding-related genes often need to determine whether a candidate gene is causally involved in microtubule dynamics, mitotic fidelity, or neuronal development. EDITGENE provides a comprehensive suite of CRISPR-based services to generate precisely engineered cell models, enabling rigorous functional validation of plus-end binding mechanisms.
Contact EDITGENE today to design your custom CRISPR model for microtubule plus-end binding research.
Related Products
| Product name | Cat.No. | Species | Gene ID | |
|---|---|---|---|---|
| TTBK2 Knockout HEK293 Cell Line | EDJ-KQ1018 | Human | 146057 | Details Get a Quote |
| CLIP1 Knockout HEK293 Cell Line | EDJ-KQ1171 | Human | 6249 | Details Get a Quote |
| STIM1 Knockout HEK293 Cell Line | EDC09869 | Human | 6786 | Details Get a Quote |
| KNSTRN Knockout HEK293 Cell Line | EDJ-KQ3313 | Human | 90417 | Details Get a Quote |
| CLIP2 Knockout HEK293 Cell Line | EDJ-KQ3816 | Human | 7461 | Details Get a Quote |
| DST Knockout HEK293 Cell Line | EDJ-KQ4147 | Human | 667 | Details Get a Quote |
| MAPRE2 Knockout HEK293 Cell Line | EDJ-KQ7234 | Human | 10982 | Details Get a Quote |
| MAPRE1 Knockout HEK293 Cell Line | EDJ-KQ7744 | Human | 22919 | Details Get a Quote |
| MAPRE3 Knockout HEK293 Cell Line | EDJ-KQ7753 | Human | 22924 | Details Get a Quote |
| CLASP2 Knockout HEK293 Cell Line | EDJ-KQ7838 | Human | 23122 | Details Get a Quote |
| CLIP3 Knockout HEK293 Cell Line | EDJ-KQ8347 | Human | 25999 | Details Get a Quote |
| CLIP4 Knockout HEK293 Cell Line | EDJ-KQ12188 | Human | 79745 | Details Get a Quote |
| CLASP1 Knockout HEK293 Cell Line | EDJ-KQ12929 | Human | 23332 | Details Get a Quote |
| APC Knockout HEK293 Cell Line | EDJ-KQ17894 | Human | 324 | Details Get a Quote |
| APC Knockout HeLa Cell Line | EDJ-KQ17952 | Human | 324 | Details Get a Quote |
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Frequently Asked Questions About microtubule plus-end binding
What is microtubule plus-end binding?
Microtubule plus-end binding (GO:0051010) is the biological process by which proteins selectively associate with the growing plus ends of microtubules, often forming comet-like structures that regulate microtubule dynamics.
What genes are involved in microtubule plus-end binding?
Key genes include MAPRE1 (EB1), MAPRE3 (EB3), CLIP1, CLASP1/2, TACC3, and SEPT9, all of which encode proteins that track microtubule plus ends.
How does EB1 regulate microtubule plus-end dynamics?
EB1 recognizes the plus end and recruits other +TIPs, and its phase separation behavior guides plus-end dynamics and mitotic spindle organization.
What diseases are linked to microtubule plus-end binding defects?
Defects are linked to cancer, neurodevelopmental disorders, and mitotic diseases such as chromosomal instability.
What methods are used to study microtubule plus-end binding?
Live-cell imaging, CRISPR knockout screens, proteomics, and in vitro reconstitution are commonly used.
Can CRISPR be used to study plus-end binding proteins?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to dissect plus-end protein functions.
What is the role of phase separation in plus-end binding?
Phase separation of EB1 and partners forms biomolecular condensates that organize plus-end dynamics and are regulated during mitosis.
How does TACC3 function at microtubule plus ends?
TACC3 regulates plus-end dynamics and cargo transport in interphase cells.
Is septin 9 involved in microtubule plus-end binding?
Yes, septin 9 modulates microtubule plus-end dynamics through its interaction with microtubules.
Why is microtubule plus-end binding important for neurons?
Plus-end tracking proteins regulate microtubule remodeling during axon outgrowth and neuronal migration.
Conclusion
Microtubule plus-end binding (GO:0051010) is a dynamic and essential biological process that orchestrates microtubule function in mitosis, intracellular transport, and neuronal development. The integration of phase separation, +TIP networks, and regulatory modifications ensures precise control of microtubule dynamics. Dysregulation of this process contributes to cancer and neurodevelopmental disorders, making it a compelling area for therapeutic targeting. CRISPR-based models and advanced imaging will continue to unravel the mechanistic details and disease relevance of plus-end binding.
References
- 1. Song X et al.. 2023. Phase separation of EB1 guides microtubule plus-end dynamics.. Nat Cell Biol 25(1):79-91 PMID: 36536176
- 2. Yang F et al.. 2024. Organization of microtubule plus-end dynamics by phase separation in mitosis.. J Mol Cell Biol 16(2) PMID: 38323478
- 3. Nakos K et al.. 2019. Regulation of microtubule plus end dynamics by septin 9.. Cytoskeleton (Hoboken) 76(1):83-91 PMID: 30144301
- 4. Mimori-Kiyosue Y. 2006. [Microtubule plus-end-binding proteins (+TIPs)].. Tanpakushitsu Kakusan Koso 51(6 Suppl):543-50 PMID: 16719310
- 5. van de Willige D et al.. 2016. Microtubule plus-end tracking proteins in neuronal development.. Cell Mol Life Sci 73(10):2053-77 PMID: 26969328
- 6. Furey C et al.. 2020. TACC3 Regulates Microtubule Plus-End Dynamics and Cargo Transport in Interphase Cells.. Cell Rep 30(1):269-283.e6 PMID: 31914393
- 7. Akhmanova A et al.. 2005. Microtubule plus-end-tracking proteins: mechanisms and functions.. Curr Opin Cell Biol 17(1):47-54 PMID: 15661518
- 8. Nehlig A et al.. 2017. Regulation of end-binding protein EB1 in the control of microtubule dynamics.. Cell Mol Life Sci 74(13):2381-2393 PMID: 28204846
