Katanin is responsible for the M-phase microtubule-severing activity in Xenopus eggs.
Microtubules are dynamic structures whose proper rearrangement during the cell cycle is essential for the positioning of membranes during interphase and for chromosome segregation during mitosis. The previous discovery of a cyclin B/cdc2-activated microtubule-severing activity in M-phase Xenopus egg extracts suggested that a microtubule-severing protein might play an important role in cell cycle-dependent changes in microtubule dynamics and organization. However, the isolation of three different microtubule-severing proteins, p56, EF1alpha, and katanin, has only confused the issue because none of these proteins is directly activated by cyclin B/cdc2. Here we use immunodepletion with antibodies specific for a vertebrate katanin homologue to demonstrate that katanin is responsible for the majority of M-phase severing activity in Xenopus eggs. This result suggests that katanin is responsible for changes in microtubules occurring at mitosis. Immunofluorescence analysis demonstrated that katanin is concentrated at a microtubule-dependent structure at mitotic spindle poles in Xenopus A6 cells and in human fibroblasts, suggesting a specific role in microtubule disassembly at spindle poles. Surprisingly, katanin was also found in adult mouse brain, indicating that katanin may have other functions distinct from its mitotic role.
McNally
F J
FJ
Section of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616, USA. fjmcnally@ucdavis.edu
Thomas
S
S
D000251:Adenosine Triphosphatases / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D000074201:Katanin; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D008938:Mitosis / Q000187:drug effects*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010063:Ovum / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000201:enzymology / Q000502:physiology; D014981:Xenopus
D000251:Adenosine Triphosphatases; D000074201:Katanin