title: Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein.
abstract: The conversion of chemical energy into mechanical force by AAA+ (ATPases associated with diverse cellular activities) ATPases is integral to cellular processes, including DNA replication, protein unfolding, cargo transport and membrane fusion. The AAA+ ATPase motor cytoplasmic dynein regulates ciliary trafficking, mitotic spindle formation and organelle transport, and dissecting its precise functions has been challenging because of its rapid timescale of action and the lack of cell-permeable, chemical modulators. Here we describe the discovery of ciliobrevins, the first specific small-molecule antagonists of cytoplasmic dynein. Ciliobrevins perturb protein trafficking within the primary cilium, leading to their malformation and Hedgehog signalling blockade. Ciliobrevins also prevent spindle pole focusing, kinetochore-microtubule attachment, melanosome aggregation and peroxisome motility in cultured cells. We further demonstrate the ability of ciliobrevins to block dynein-dependent microtubule gliding and ATPase activity in vitro. Ciliobrevins therefore will be useful reagents for studying cellular processes that require this microtubule motor and may guide the development of additional AAA+ ATPase superfamily inhibitors.
authors.affiliation: Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California 94305, USA.
authors.forename: Ari J
authors.initials: AJ
authors.lastname: Firestone
authors.forename: Joshua S
authors.initials: JS
authors.lastname: Weinger
authors.forename: Maria
authors.initials: M
authors.lastname: Maldonado
authors.forename: Kari
authors.initials: K
authors.lastname: Barlan
authors.forename: Lance D
authors.initials: LD
authors.lastname: Langston
authors.forename: Michael
authors.initials: M
authors.lastname: O'Donnell
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Gelfand
authors.forename: Tarun M
authors.initials: TM
authors.lastname: Kapoor
authors.forename: James K
authors.initials: JK
authors.lastname: Chen
mesh_terms: D000818:Animals; D002417:Cattle; D002923:Cilia / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003593:Cytoplasm / Q000201:enzymology*; D056445:Cytoplasmic Dyneins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D053823:Hedgehog Proteins / Q000378:metabolism; D018386:Kinetochores / Q000187:drug effects / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism; D020460:Melanosomes / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D008970:Molecular Weight; D009068:Movement / Q000187:drug effects; D041681:NIH 3T3 Cells; D020675:Peroxisomes / Q000187:drug effects / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects; D052999:Quinazolinones / Q000737:chemistry* / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology
chemical_list: D004791:Enzyme Inhibitors; D053823:Hedgehog Proteins; D051741:Kruppel-Like Transcription Factors; D052999:Quinazolinones; D056445:Cytoplasmic Dyneins