Key Points
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Microtubule organization and dynamics are controlled by proteins that associate with the two microtubule extremities, the plus end and the minus end.
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Proteins accumulating at microtubule ends can promote or inhibit microtubule polymerization, enhance or block microtubule disassembly, or induce transitions between microtubule growth and shortening.
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Microtubule plus-end-tracking proteins (+TIPs) can accumulate at microtubule ends by recognizing the stabilizing GTP cap at growing microtubule tips or the curvature of the outermost part of the microtubule, or by plus-end-directed motor activity. This accumulation can be enhanced by electrostatic interactions between positively charged protein domains and the negatively charged microtubule lattice.
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+TIPs form extensive interaction networks, which depend on a limited number of protein motifs and modules that bind to each other with moderate affinity, allowing rapid remodelling of the end-associated complexes during microtubule growth and shortening. The recruitment of proteins with SxIP and cytoskeleton-associated protein Gly-rich (CAP-Gly) domains by the 'autonomous' +TIPs of the end-binding protein (EB) family plays a major part in the formation of these networks.
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+TIP networks are responsible for a large range of cellular functions, such as microtubule guidance along other cytoskeletal elements, microtubule attachment to the cell cortex, kinetochores and intracellular membrane organelles, positioning of microtubule arrays and signalling.
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Microtubule minus-end-targeting proteins (−TIPs) of the calmodulin-regulated spectrin-associated protein (CAMSAP) and Patronin family accumulate at free, growing microtubule minus ends and control the architecture of microtubule networks by stabilizing non-centrosomal microtubules.
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A wide range of diverse pharmacological agents can target microtubule tips either directly or indirectly and cooperate with +TIPs in regulating the dynamics of microtubule ends.
Abstract
Microtubules have fundamental roles in many essential biological processes, including cell division and intracellular transport. They assemble and disassemble from their two ends, denoted the plus end and the minus end. Significant advances have been made in our understanding of microtubule plus-end-tracking proteins (+TIPs) such as end-binding protein 1 (EB1), XMAP215, selected kinesins and dynein. By contrast, information on microtubule minus-end-targeting proteins (−TIPs), such as the calmodulin-regulated spectrin-associated proteins (CAMSAPs) and Patronin, has only recently started to emerge. Here, we review our current knowledge of factors, including microtubule-targeting agents, that associate with microtubule ends to control the dynamics and function of microtubules during the cell cycle and development.
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Acknowledgements
The authors are indebted to Natacha Olieric for help preparing the illustrations, to Shasha Hua and Renu Mohan for preparing original fluorescence images. A.A. is supported by a European Research Council (ERC) Synergy grant and grants from the Netherlands Organization for Scientific Research, and M.O.S. is supported by grants from the Swiss National Science Foundation and the Swiss SystemsX.ch initiative.
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FURTHER INFORMATION
Glossary
- Neurons
-
Cells that process and transmit information through electrical and chemical signals.
- Protofilaments
-
Straight rows of longitudinally aligned tubulin dimers in microtubules.
- Microtubule plus end
-
The dynamic end of a microtubule, which alternates in vivo between periods of growth and shrinkage and is often directed towards the cell surface. Microtubule plus ends grow quickly in vitro.
- Microtubule minus end
-
The less-dynamic end of a microtubule. Microtubule minus ends do grow, albeit slowly. In cells, they are often stabilized (for example, by attachment to the centrosome) or function as sites of microtubule depolymerization. Microtubule minus ends grow slowly in vitro.
- Catastrophe
-
The transition of a microtubule from growth to shortening.
- Rescues
-
Transitions of microtubules from shortening to growth.
- GTP islands
-
GTP-bound tubulin dimer patches in the microtubule shaft.
- Kinesin
-
A microtubule-based molecular motor, often directed towards the plus ends of microtubules.
- Motor processivity
-
The ability of motors to move long distances along a cytoskeletal filament without dissociation.
- Axonemal microtubules
-
The central components of axonemes. Numerous eukaryotic cells carry whip-like appendages (cilia or eukaryotic flagella), the inner cores of which consist of a microtubule-based structure called the axoneme. The axoneme contains axonemal microtubules, which function as the 'skeleton' of these cell protrusions, giving them support, enabling transport and, in some cases, causing bending motion.
- Kinetochores
-
Specialized regions on chromosomes that are connected to microtubules and motor proteins during cell division in eukaryotes. Kinetochores function in the separation of chromosome pairs.
- Cytoskeleton-associated protein Gly-rich domain
-
(CAP-Gly domain). An ∼70-residue protein domain that is characterized by several Gly residues, which are involved in shaping the loop regions of the globular fold. CAP-Gly domains contain a unique hydrophobic cavity that encompasses the highly conserved GKNDG sequence motif responsible for targeting CAP-Gly domains to the carboxy-terminal EEY/F motifs of γ-tubulin, end-binding protein, cytoplasmic linker protein 170 and SLAIN.
- Dynein
-
A large, minus-end-directed, multisubunit microtubule motor protein that is involved in several cellular processes, including cell division, migration and intracellular transport.
- Calponin homology domain
-
(CH domain). An ∼100-residue actin- or microtubule-binding domain that is common to many actin-binding proteins, including cytoskeletal and signalling proteins, and microtubule-associated proteins such as end-binding proteins, calponin-homology and microtubule-associated protein (CLAMP) and highly expressed in cancer protein 1 (HEC1).
- Coiled-coil domain
-
A protein structural motif that mediates subunit oligomerization. Coiled-coils contain between two and five α-helices that twist around each other to form a supercoil.
- Carboxy-terminal EEY/F motif
-
A highly specific and conserved sequence motif found at the C termini of α-tubulin, end-binding proteins, cytoplasmic linker protein 170 and SLAIN. The EEY/F motif is the target of cytoskeleton-associated protein Gly-rich domains.
- EB homology domain
-
(EBH domain). An ∼50-amino-acid domain found at the carboxyl termini of end-binding proteins. It constitutes of a pair of helix–loop–helix segments forming an antiparallel four-helix bundle. The highly conserved residues of the EBH domain form a surface patch that contains a deep hydrophobic cavity, which serves as an interaction site for binding partners.
- TOG domains
-
HEAT (Huntingtin, elongation factor 3, protein phosphatase 2A and target of rapamycin1) repeat-containing tubulin-binding domains named after their discovery in the human microtubule-associated protein ch-TOG (colonic and hepatic tumour-overexpressed gene).
- Doublecortin domains
-
(DC domains). Regions found in the amino terminus of the protein doublecortin that consist of tandem repeated copies of an approximately 80-amino-acid sequence region.
- Nuclear division cycle 80 complex
-
(NDC80 complex). A multisubunit protein complex that mediates the attachment of microtubule plus ends to kinetochores.
- DUO1 and MPS1-interacting 1 complex
-
(DAM1 complex). A multisubunit protein complex (also known as the DASH complex) that is involved in kinetochore–microtubule attachments in yeast and can form rings around microtubules. It may provide a dynamic linkage at depolymerizing or polymerizing microtubule ends for force generation.
- SxIP motif
-
An amino acid sequence motif of Ser-any amino acid-Ile-Pro, embedded in an intrinsically disordered positively charged sequence region that is found in many microtubule plus-end-tracking proteins. The SxIP motif is specifically recognized by the EB homology domain of EBs.
- Kinases
-
Enzymes that catalyse the transfer of phosphate groups from ATP to specific substrates, such as particular amino acid residues in proteins. This process is termed phosphorylation.
- Actin
-
A protein that forms microfilaments in most cells. Actin filaments represent one of the three cytoskeletal filament systems besides microtubules and intermediate filaments.
- Myosin V
-
A subclass of the myosin family of actin-dependent motor proteins that is required for the transport of vesicles or mRNA cargo.
- Phosphoinositide
-
A phosphorylated form of phosphatidylinositol, a member of a family of lipids that has important roles in lipid signalling, cell signalling and membrane trafficking.
- Focal adhesions
-
Integrin-mediated cell–substrate adhesion structures that anchor the ends of actin filaments (also known as stress fibres) and mediate strong attachments to substrates. They also function as integrin signalling platforms.
- Neuromuscular junctions
-
Structures that connect the nervous system to the muscular system through synapses between nerve fibres and muscle cells. In vertebrates, the small-molecule neurotransmitter acetylcholine is released by the neuron and binds acetylcholine receptors in the plasma membrane of the muscle cell.
- Retrograde transport
-
The movement of molecules or organelles inwards, away from the plasma membrane and towards the cell body. Often mediated by the microtubule minus-end-directed motor dynein.
- Guanine nucleotide exchange factors
-
(GEFs). Proteins that facilitate the exchange of GDP for GTP in the nucleotide-binding pocket of GTP-binding proteins.
- Neurites
-
Projections extending from the cell body of a neuronal cell. In differentiated neurons, neurites are distinguished into axons (projections that transmit signals) and dendrites (projections that receive signals).
- Endoplasmic reticulum
-
(ER). A membrane organelle in eukaryotic cells that has important functions in protein synthesis, folding and processing, as well as in lipid and sugar metabolism and calcium storage. It forms an interconnected network of flattened, membrane-enclosed sacs or tubes.
- γ-tubulin ring complex
-
(γ-TURC). A multisubunit protein complex that binds to microtubule minus ends and nucleates microtubules, for example, at the centrosome. The complex contains a special isoform of tubulin, γ-tubulin.
- Centrosomes
-
Organelles that form the main microtubule-organizing centre (MTOC) of most animal cells. They consist of cylindrical, microtubule-based structures called centrioles and their surrounding material, which can nucleate and anchor microtubules.
- CKK domain
-
(Calmodulin-regulated spectrin-associated protein (CAMSAP), KIAA1078 and KIAA1543 domain). A domain that occurs at the carboxyl termini of the CAMSAPs and Patronin. CKK domains are thought to bind microtubules.
- Poleward microtubule flux
-
The process of translocation of spindle microtubules towards spindle poles, coupled to the depolymerization of their minus ends.
- Peptide aptamers
-
Small proteins containing a 10–25-amino-acid peptide that binds with high specificity and affinity to a molecular target.
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Akhmanova, A., Steinmetz, M. Control of microtubule organization and dynamics: two ends in the limelight. Nat Rev Mol Cell Biol 16, 711–726 (2015). https://doi.org/10.1038/nrm4084
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DOI: https://doi.org/10.1038/nrm4084
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