title: Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport.
abstract: Polarized transport in neurons is fundamental for the formation of neuronal circuitry. A motor domain-containing truncated KIF5 (a kinesin-1) recognizes axonal microtubules, which are enriched in EB1 binding sites, and selectively accumulates at the tips of axons. However, it remains unknown what cue KIF5 recognizes to result in this selective accumulation. We found that axonal microtubules were preferentially stained by the anti-GTP-tubulin antibody hMB11. Super-resolution microscopy combined with EM immunocytochemistry revealed that hMB11 was localized at KIF5 attachment sites. In addition, EB1, which binds preferentially to guanylyl-methylene-diphosphate (GMPCPP) microtubules in vitro, recognized hMB11 binding sites on axonal microtubules. Further, expression of hMB11 antibody in neurons disrupted the selective accumulation of truncated KIF5 in the axon tips. In vitro studies revealed approximately threefold stronger binding of KIF5 motor head to GMPCPP microtubules than to GDP microtubules. Collectively, these data suggest that the abundance of GTP-tubulin in axonal microtubules may underlie selective KIF5 localization and polarized axonal vesicular transport.
authors.affiliation: Department of Cell Biology and Anatomy, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Tokyo 113-0033, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Nakata
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Niwa
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okada
authors.forename: Franck
authors.initials: F
authors.lastname: Perez
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Hirokawa
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001369:Axons / Q000378:metabolism; D001692:Biological Transport; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000276:immunology / Q000378:metabolism*; D013552:Swine; D014404:Tubulin / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D008869:Microtubule-Associated Proteins; D014404:Tubulin; C004805:5'-guanylylmethylenebisphosphonate; D006160:Guanosine Triphosphate; D016547:Kinesins