Force measurements on cargoes in living cells reveal collective dynamics of microtubule motors.

Abstract:

:Many cellular cargoes move bidirectionally along microtubules, driven by teams of plus- and minus-end-directed motor proteins. To probe the forces exerted on cargoes during intracellular transport, we examined latex beads phagocytosed into living mammalian macrophages. These latex bead compartments (LBCs) are encased in membrane and transported along the cytoskeleton by a complement of endogenous kinesin-1, kinesin-2, and dynein motors. The size and refractive index of LBCs makes them well-suited for manipulation with an optical trap. We developed methods that provide in situ calibration of the optical trap in the complex cellular environment, taking into account any variations among cargoes and local viscoelastic properties of the cytoplasm. We found that centrally and peripherally directed forces exerted on LBCs are of similar magnitude, with maximum forces of ~20 pN. During force events greater than 10 pN, we often observe 8-nm steps in both directions, indicating that the stepping of multiple motors is correlated. These observations suggest bidirectional transport of LBCs is driven by opposing teams of stably bound motors that operate near force balance.

authors

Hendricks AG,Holzbaur EL,Goldman YE

doi

10.1073/pnas.1215462109

subject

Has Abstract

pub_date

2012-11-06 00:00:00

pages

18447-52

issue

45

eissn

0027-8424

issn

1091-6490

pii

1215462109

journal_volume

109

pub_type

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