Mechanical coupling of microtubule-dependent motor teams during peroxisome transport in Drosophila S2 cells

Abstract

Background Intracellular transport requires molecular motors that step along cytoskeletal filaments actively dragging cargoes through the crowded cytoplasm. Here, we explore the interplay of the opposed polarity motors kinesin-1 and cytoplasmic dynein during peroxisome transport along microtubules in Drosophila S2 cells. Methods We used single particle tracking with nanometer accuracy and millisecond time resolution to extract quantitative information on the bidirectional motion of organelles. The transport performance was studied in cells expressing a slow chimeric plus-end directed motor or the kinesin heavy chain. We also analyzed the influence of peroxisomes membrane fluidity in methyl-β-ciclodextrin treated cells. The experimental data was also confronted with numerical simulations of two well-established tug of war scenarios. Results and conclusions The velocity distributions of retrograde and anterograde peroxisomes showed a multi-modal pattern suggesting that multiple motor teams drive transport in either direction. The chimeric motors interfered with the performance of anterograde transport and also reduced the speed of the slowest retrograde team. In addition, increasing the fluidity of peroxisomes membrane decreased the speed of the slowest ante-rograde and retrograde teams. General significance Our results support the existence of a crosstalk between opposed-polarity motor teams. Moreover, the slowest teams seem to mechanically communicate with each other through the membrane to trigger transport.

Retraction of rod-like mitochondria during microtubule-dependent transport

“…Rod-shaped mitochondria are attractive to explore the performance of motor-teams since their elongated shape determines a broad distribution of motors on the surface. As we mentioned before, the mechanical communication amongst active motors is expected to be different from that observed in small, rounded organelles as peroxisomes [ 8 ] where active motors are spatially confined to a smaller area and thus they are probably closer to each other.…”

“…The team performance depends on the biophysical properties of the motors (e.g. [ 3–5 ]), the number of motors participating in the team [ 6 , 7 ], and their mechanical coupling when attached to the cargo surface [ 8–10 ] amongst other factors.…”

The Generation of Dynein Networks by Multi-Layered Regulation and Their Implication in Cell Division

“…Multiple dyneins are connected directly or indirectly in the cell through molecular or physical interactions to form a functional network of dyneins. For example, intracellular vesicles usually contain multiple dyneins (Hendricks et al, 2010;Encalada et al, 2011;Rai et al, 2013Rai et al, , 2016De Rossi et al, 2017;Chowdary et al, 2018;Cella Zanacchi et al, 2019), and the clustering enables a team of dyneins to produce collective large stall forces Rai et al, 2013). A recent super-resolution study also reported that dynein forms nanoclusters composed of up to seven dimers on microtubules (Cella Zanacchi et al, 2019).…”

Stable tug-of-war between kinesin-1 and cytoplasmic dynein upon different ATP and roadblock concentrations

“…Besides activation by adaptor proteins, dynein can also be mechanically activated (Monzon and Scharrel et al, 2019;Torisawa et al, 2014). Ally et al (2009) have shown that a mechanical motor coupling is needed for cargo transport and De Rossi et al (2017) hypothesize that motors might activate each other by exerting forces on the opposite motor team. Mechanical dynein activation has been shown to determine the velocity in unidirectional dynein-driven transport (Monzon and Scharrel et al, 2019).…”