In vivo optical trapping indicates kinesin’s stall force is reduced by dynein during intracellular transport

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 110; no. 9; pp. 3381 - 3386
• Main Authors: Blehm, Benjamin H., Schroer, Trina A., Trybus, Kathleen M., Chemla, Yann R., Selvin, Paul R.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 26.02.2013; National Acad Sciences
• Subjects: Animals; Biological Sciences; Biological Transport; Biomechanical Phenomena - physiology; Calibration; Cells; Cytoplasm; Data acquisition; Dictyostelium - cytology; Dictyostelium - metabolism; dynein ATPase; Dyneins - metabolism; Freight; Histograms; Humans; Intracellular Space - metabolism; kinesin; Kinesin - metabolism; Lasers; Microtubules; Models, Biological; Molecules; Motors; optical traps; Optical Tweezers; Organelles; Protein Binding; Stall; Stiffness
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1219961110

Kinesin and dynein are fundamental components of intracellular transport, but their interactions when simultaneously present on cargos are unknown. We built an optical trap that can be calibrated in vivo during data acquisition for each individual cargo to measure forces in living cells. Comparing directional stall forces in vivo and in vitro, we found evidence that cytoplasmic dynein is active during minus- and plus-end directed motion, whereas kinesin is only active in the plus direction. In vivo, we found outward (∼plus-end) stall forces range from 2 to 7 pN, which is significantly less than the 5- to 7-pN stall force measured in vitro for single kinesin molecules. In vitro measurements on beads with kinesin-1 and dynein bound revealed a similar distribution, implying that an interaction between opposite polarity motors causes this difference. Finally, inward (∼minus-end) stalls in vivo were 2–3 pN, which is higher than the 1.1-pN stall force of a single dynein, implying multiple active dynein.