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Cargo adaptors regulate stepping and force generation of mammalian dynein-dynactin

Elshenawy, Mohamed M ; Canty, John T ; Oster, Liya ; Ferro, Luke S ; Zhou, Zhou ; Blanchard, Scott C ; Yildiz, Ahmet

Nature chemical biology, 2019-11, Vol.15 (11), p.1093-1101 [Periódico revisado por pares]

United States: Nature Publishing Group

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  • Título:
    Cargo adaptors regulate stepping and force generation of mammalian dynein-dynactin
  • Autor: Elshenawy, Mohamed M ; Canty, John T ; Oster, Liya ; Ferro, Luke S ; Zhou, Zhou ; Blanchard, Scott C ; Yildiz, Ahmet
  • Assuntos: Adapters ; Animals ; Cargo ; Conformation ; Dynactin ; Dynactin Complex - chemistry ; Dynactin Complex - metabolism ; Dynein ; Dyneins - chemistry ; Dyneins - metabolism ; Humans ; Kinesin ; Load distribution ; Microtubules ; Motility ; Protein Binding ; Velocity
  • É parte de: Nature chemical biology, 2019-11, Vol.15 (11), p.1093-1101
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
    M.M.E., J.C., and A.Y. conceived the study and designed the experiments. M.M.E., J.C., and L.F. prepared the constructs and isolated the proteins. Z.Z. and S.B. synthesized the fluorescent dyes. M.M.E. labeled the proteins with DNA and fluorescent dyes and performed the fluorescence motility experiments. J.C. performed bulk ATPase and the MT bridge assays. J.C. and L.O. performed fluorescent tracking assays. M.M.E. performed optical-trapping assays. M.M.E., J.C., and A.Y. wrote the manuscript, and all authors read and commented on the manuscript.
    Author contributions
  • Descrição: Cytoplasmic dynein is an ATP-driven motor that transports intracellular cargos along microtubules. Dynein adopts an inactive conformation when not attached to a cargo, and motility is activated when dynein assembles with dynactin and a cargo adaptor. It was unclear how active dynein-dynactin complexes step along microtubules and transport cargos under tension. Using single-molecule imaging, we showed that dynein-dynactin advances by taking 8 to 32-nm steps toward the microtubule minus end with frequent sideways and backward steps. Multiple dyneins collectively bear a large amount of tension because the backward stepping rate of dynein is insensitive to load. Recruitment of two dyneins to dynactin increases the force generation and the likelihood of winning against kinesin in a tug-of-war but does not directly affect velocity. Instead, velocity is determined by cargo adaptors and tail-tail interactions between two closely packed dyneins. Our results show that cargo adaptors modulate dynein motility and force generation for a wide range of cellular functions.
  • Editor: United States: Nature Publishing Group
  • Idioma: Inglês
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