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

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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.

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Fig. 1: DDR hydrolyzes ATP and moves faster than DDB.
Fig. 2: The stepping behavior of mammalian dynein–dynactin in unloaded conditions.
Fig. 3: Velocity and stepping of mammalian dynein–dynactin under load.
Fig. 4: Tail–tail interactions increase the velocity of dynein–dynactin.
Fig. 5: Dynein–dynactin multiplicity increases total force generation.
Fig. 6: Recruitment of a second dynein shifts the balance toward the MT minus end in a tug-of-war.

Data availability

All data that support the conclusions are available from the authors on request.

Code availability

Code used in this paper is available from the corresponding author upon request.


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We are grateful to the members of the Yildiz laboratory for helpful discussions, A. P. Carter, L. Urnavicius, and S. Lacey (MRC, Cambridge) for providing plasmids and helping with protein expression and purification, and D. Drubin and C. Kaplan for multicolor TIRF microscopy. This work was funded by grants from the NIH (GM094522), and NSF (MCB-1055017, MCB-1617028) to A.Y., a grant from the NIH (GM098859) to S.C.B. and NIH F31 fellowship to L.F.

Author information

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 MT bridge assays. J.C. and L.O. performed fluorescence tracking assays. M.M.E. performed optical trapping assays. M.M.E., J.C. and A.Y. wrote the manuscript. All authors read and commented on the manuscript.

Correspondence to Ahmet Yildiz.

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S.C.B. holds an equity interest in Lumidyne Technologies. All other authors declare no competing interests.

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Supplementary information

Supplementary Information

Supplementary Tables 1–3 and Supplementary Figures 1–8

Reporting Summary

Supplementary Video 1

Processive motility of single DDB and DDR complexes along MTs in 1 mM ATP.

Supplementary Video 2

Helical movement of DDB- and DDR-driven beads along an MT bridge.

Supplementary Video 3

Linking of two DDBs via DNA hybridization.

Supplementary Video 4

Motility of DDB–kinesin and DDR–kinesin colocalizers.

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Elshenawy, M.M., Canty, J.T., Oster, L. et al. Cargo adaptors regulate stepping and force generation of mammalian dynein–dynactin. Nat Chem Biol 15, 1093–1101 (2019) doi:10.1038/s41589-019-0352-0

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