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

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 complex...

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Published inNature chemical biology Vol. 15; no. 11; pp. 1093 - 1101
Main Authors Elshenawy, Mohamed M., Canty, John T., Oster, Liya, Ferro, Luke S., Zhou, Zhou, Blanchard, Scott C., Yildiz, Ahmet
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.11.2019
Nature Publishing Group
Subjects
631/1647/245
631/337/2265
631/80
631/92/56
Adapters
Animals
Biochemical Engineering
Biochemistry
Bioorganic Chemistry
Cargo
Cell Biology
Chemistry
Chemistry and Materials Science
Chemistry/Food Science
Conformation
Dynactin
Dynactin Complex - chemistry
Dynactin Complex - metabolism
Dynein
Dyneins - chemistry
Dyneins - metabolism
Humans
Kinesin
Load distribution
Microtubules
Motility
Protein Binding
Velocity
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Summary: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. Single-molecule analysis revealed that the velocity and force generation of the mammalian dynein–dynactin complex is regulated by activating adaptors and tail–tail interactions between two dyneins.
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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
ISSN:1552-4450
1552-4469
DOI:10.1038/s41589-019-0352-0