title: Cytoplasmic Dynein Antagonists with Improved Potency and Isoform Selectivity.
abstract: Cytoplasmic dyneins 1 and 2 are related members of the AAA+ superfamily (ATPases associated with diverse cellular activities) that function as the predominant minus-end-directed microtubule motors in eukaryotic cells. Dynein 1 controls mitotic spindle assembly, organelle movement, axonal transport, and other cytosolic, microtubule-guided processes, whereas dynein 2 mediates retrograde trafficking within motile and primary cilia. Small-molecule inhibitors are important tools for investigating motor protein-dependent mechanisms, and ciliobrevins were recently discovered as the first dynein-specific chemical antagonists. Here, we demonstrate that ciliobrevins directly target the heavy chains of both dynein isoforms and explore the structure-activity landscape of these inhibitors in vitro and in cells. In addition to identifying chemical motifs that are essential for dynein blockade, we have discovered analogs with increased potency and dynein 2 selectivity. These antagonists effectively disrupt Hedgehog signaling, intraflagellar transport, and ciliogenesis, making them useful probes of these and other cytoplasmic dynein 2-dependent cellular processes.
authors.forename: Stephanie K
authors.initials: SK
authors.lastname: See
authors.forename: Sascha
authors.initials: S
authors.lastname: Hoogendoorn
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Chung
authors.forename: Fan
authors.initials: F
authors.lastname: Ye
authors.affiliation: Laboratory of Chemistry and Cell Biology, Rockefeller University , New York City, New York 10065, United States.
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Steinman
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Sakata-Kato
authors.affiliation: Laboratory of Chemistry and Cell Biology, Rockefeller University , New York City, New York 10065, United States.
authors.forename: Rand M
authors.initials: RM
authors.lastname: Miller
authors.affiliation: Laboratory of Chemistry and Cell Biology, Rockefeller University , New York City, New York 10065, United States.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Cupido
authors.affiliation: Medical Research Council Laboratory of Molecular Biology , Division of Structural Studies, Cambridge CB2 0QH, United Kingdom.
authors.forename: Ruta
authors.initials: R
authors.lastname: Zalyte
authors.affiliation: Medical Research Council Laboratory of Molecular Biology , Division of Structural Studies, Cambridge CB2 0QH, United Kingdom.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Carter
authors.forename: Maxence V
authors.initials: MV
authors.lastname: Nachury
authors.affiliation: Laboratory of Chemistry and Cell Biology, Rockefeller University , New York City, New York 10065, United States.
authors.forename: Tarun M
authors.initials: TM
authors.lastname: Kapoor
authors.forename: James K
authors.initials: JK
authors.lastname: Chen
mesh_terms: D000818:Animals; D056445:Cytoplasmic Dyneins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D053823:Hedgehog Proteins / Q000502:physiology; D051379:Mice; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000737:chemistry; D052999:Quinazolinones / Q000737:chemistry / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D053823:Hedgehog Proteins; D020033:Protein Isoforms; D052999:Quinazolinones; D056445:Cytoplasmic Dyneins