Katanin is a microtubule-severing AAA protein. It is named after the Japanese sword called a katana. Katanin is a heterodimeric protein first discovered in sea urchins. It contains a 60 kDa ATPase subunit, encoded by KATNA1, which functions to sever microtubules. This subunit requires ATP and the presence of microtubules for activation. The second 80 kDA subunit, encoded by KATNB1, regulates the activity of the ATPase and localizes the protein to centrosomes. Electron microscopy shows that katanin forms 14–16 nm rings in its active oligomerized state on the walls of microtubules (although not around the microtubule).
Mechanism and regulation of microtubule length Structural analysis using electron microscopy has revealed that microtubule protofilaments change from a straight to a curved conformation upon GTP hydrolysis of β-tubulin. However, when these protofilaments are part of a polymerized microtubule, the stabilizing interactions created by the surrounding lattice lock subunits into a straight conformation, even after GTP hydrolysis. In order to disrupt these stable interactions, katanin, once bound to ATP, oligomerizes into a ring structure on the microtubule wall - in some cases oligomerization increases the affinity of katanin for microtubules and stimulates its ATPase activity. Once this structure is formed, katanin hydrolyzes ATP, and likely undergoes a conformational change that puts mechanical strain on the tubulin subunits, which destabilizes their interactions within the microtubule lattice. The predicted conformational change also likely decreases the affinity of katanin for tubulin as well as for other katanin proteins, which leads to disassembly of the katanin ring structure, and recycling of the individual inactivated proteins. The severing of microtubules by katanin is regulated by protective microtubule-associated proteins (MAPs), and the p80 subunit (p60 severs microtubules much better in the presence of p80). These mechanisms have different consequences, depending on where in the cell they are activated or disrupted. For example, allowing katanin-mediated severing at the centrosome releases microtubules for free movement. In one experiment, anti-katanin antibodies were injected into a cell, causing a large accumulation of microtubules around the centrosome and inhibition of microtubule outgrowth. Therefore, katanin-mediated severing may serve to maintain organization in the cytoplasm by promoting microtubule disassembly and efficient movement. During cell division, severing at the spindle pole produces free microtubule ends and allows poleward flux of tubulin and retraction of the microtubule. Severing microtubules in the cytoplasm facilitates treadmilling and mobility, which is important during development.
Role in cell division Katanin-mediated microtubule severing is an important step in mitosis and meiosis. It has been shown that katanin is responsible for severing microtubules during M-phase in Xenopus laevis. The disassembly of microtubules from their interphase structures is necessary to prepare the cell and the mitotic spindle for cell division. This regulation is indirect: MAP proteins, which protect the microtubules from being severed during interphase, dissociate and allow katanin to act. In addition, katanin is responsible for severing microtubules at the mitotic spindles when disassembly is required to segregate sister chromatids during anaphase. Similar results have been obtained in relation to katanin's activity during meiosis in C. elegans. It was reported that Mei-1 and Mei-2 to encode similar proteins to the p60 and p80 subunits of katanin. Using antibodies, these two proteins were found to localize at the ends of microtubules in the meiotic spindle, and, when expressed in HeLa cells, these proteins initiated microtubule severing. These findings indicate that katanin serves a similar purpose in both mitosis and meiosis in segregating chromatids toward the spindle poles.
Role in development Katanin is important in the development of many organisms. Both elimination and overexpression of katanin is deleterious to axonal growth, and, thus, katanin must be carefully regulated for proper neural development. In particular, severing microtubules in specific cellular spaces allows fragments to test various routes of growth. Katanin has proved necessary in this task. An experiment using time-lapse digital imaging of fluorescently labeled tubulin demonstrated that axon growth cones pause, and microtubules fragment, at sites of branching during neural development. A similar experiment using fluorescently labeled tubulin observed local microtubule fragmentation in newt lung cell lamellipodia during developmental migration, in which the fragments run perpendicular to the advancing cell membrane to aid exploration. The local nature of both fragmentation events likely indicates regulation by katanin because it can be concentrated in specific cellular regions. This is supported by a study that demonstrated that the Fra2 mutation, which affects a katanin orthologue in Arabidopsis thaliana, leads to an aberrant disposition of cellulose microfibrils along the developing cell wall in these plants. This mutation produced a phenotype with reduced cell elongation, which suggests katanin's significance in development across a wide range of organisms.
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