title: Two domains of p80 katanin regulate microtubule severing and spindle pole targeting by p60 katanin.
abstract: The assembly and function of the mitotic spindle requires the activity of a number of microtubule-binding proteins. Some microtubule-binding proteins bind microtubules in vitro but do not co-localize with microtubules in interphase cells. Instead these proteins associate with specific subregions of the mitotic spindle. Katanin, a heterodimeric microtubule-severing ATPase, is found localized at mitotic spindle poles. In this paper we demonstrate that human p60 katanin and the C-terminal domain of human p80 katanin both bind microtubules in vitro. Association of these two proteins results in an increased microtubule affinity and increased microtubule-severing activity in vitro. Association of these subunits in transfected HeLa cells increases microtubule disassembly activity and targeting to spindle poles. The N-terminal WD40 domain of p80 katanin acts as a negative regulator of microtubule disassembly activity and is also required for spindle pole localization, possibly through interactions with another spindle-pole protein. These results support a model in which katanin is targeted to spindle poles through a combination of direct microtubule binding by the p60 subunit and through interactions between the WD40 domain and an unknown protein. We propose that both domains of p80 are essential in precisely regulating katanin's activity in vivo.
authors.affiliation: Section of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA. fjmcnally@ucdavis.edu
authors.forename: K P
authors.initials: KP
authors.lastname: McNally
authors.forename: O A
authors.initials: OA
authors.lastname: Bazirgan
authors.forename: F J
authors.initials: FJ
authors.lastname: McNally
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D006367:HeLa Cells; D006801:Humans; D000074201:Katanin; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D008941:Spindle Apparatus / Q000378:metabolism*
chemical_list: D000251:Adenosine Triphosphatases; D000074201:Katanin