Dynein Clusters into Lipid Microdomains on Phagosomes to Drive Rapid Transport toward Lysosomes

Rai, Ashim; Pathak, Divya; Thakur, Satbir; Singh, Shampa; Dubey, Alok Kumar; Mallik, Roop

doi:10.1016/j.cell.2015.12.054

Cited by 195 publications

(330 citation statements)

References 46 publications

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“…Conceivably then, reggies might organize microdomains with specific lipids and lipid modified proteins and promote their vesicular transport and delivery to specific regions such as the PSD. This complies with a recent report showing that dynein clusters in reggie/flotillin microdomains during transport and maturation of phagosomes to lysosomes in Leishmania donovani (Rai et al, 2016).…”

Section: Reggies In Raftssupporting

confidence: 79%

Reggie-1 and reggie-2 (flotillins) participate in Rab11a-dependent cargo trafficking, spine synapse formation and LTP-related AMPA receptor (GluA1) surface exposure in mouse hippocampal neurons

Bodrikov

Pauschert

Kochlamazashvili

et al. 2017

Experimental Neurology

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a b s t r a c tReggie-1 and -2 (flotillins) reside at recycling vesicles and promote jointly with Rab11a the targeted delivery of cargo. Recycling is essential for synapse formation suggesting that reggies and Rab11a may regulate the development of spine synapses. Recycling vesicles provide cargo for dendritic growth and recycle surface glutamate receptors (AMPAR, GluA) for long-term potentiation (LTP) induced surface exposure. Here, we show reduced number of spine synapses and impairment of an in vitro correlate of LTP in hippocampal neurons from reggie-1 k.o. (Flot2−/−) mice maturating in culture. These defects apparently result from reduced trafficking of PSD-95 revealed by live imaging of 10 div reggie-1 k.o. (Flot2 −/−) neurons and likely impairs co-transport of cargo destined for spines: N-cadherin and the glutamate receptors GluA1 and GluN1. Impaired cargo trafficking and fewer synapses also emerged in reggie-1 siRNA, reggie-2 siRNA, and reggie-1 and -2 siRNA-treated neurons and was in siRNA and k.o. neurons rescued by reggie-1-EGFP and CA-Rab11a-EGFP. While correlative expressional changes of specific synapse proteins were observed in reggie-1 k.o. (Flot2−/−) brains in vivo, this did not occur in neurons maturating in vitro. Our work suggests that reggie-1 and reggie-2 function at Rab11a recycling containers in the transport of PSD-95, N-cadherin, GluA1 and GluN1, and promote (together with significant signaling molecules) spine-directed trafficking, spine synapse formation and the in vitro correlate of LTP.

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Section: Reggies In Raftssupporting

confidence: 79%

Reggie-1 and reggie-2 (flotillins) participate in Rab11a-dependent cargo trafficking, spine synapse formation and LTP-related AMPA receptor (GluA1) surface exposure in mouse hippocampal neurons

Bodrikov

Pauschert

Kochlamazashvili

et al. 2017

Experimental Neurology

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“…Nelson et al [63] demonstrated that the fluidity of the bilayer affects the velocity of myosin teams synthetically anchored to phospholipid vesicles. On the other hand, dyneins cluster into raft-like microdomains on phagosomes improving their performance as a team [72]. These previous works show that the cargo membrane plays a relevant role on the performance of motor teams.…”

Section: Resultsmentioning

confidence: 99%

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

Rossi

Wetzler

Benseñor

et al. 2017

Biochimica et Biophysica Acta (BBA) - General Subjects

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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.

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“…This results in less slippage of the motors and thus an increased transport efficiency. Along these lines, a recent study has reported that cytoplasmic dynein clusters into lipid microdomains on phagosomes to drive rapid transport toward lysosomes (41).…”

Section: Discussionmentioning

confidence: 99%

Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity

Grover

Fischer

Schwarz

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

111

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In eukaryotic cells, membranous vesicles and organelles are transported by ensembles of motor proteins. These motors, such as kinesin-1, have been well characterized in vitro as single molecules or as ensembles rigidly attached to nonbiological substrates. However, the collective transport by membrane-anchored motors, that is, motors attached to a fluid lipid bilayer, is poorly understood. Here, we investigate the influence of motors' anchorage to a lipid bilayer on the collective transport characteristics. We reconstituted "membrane-anchored" gliding motility assays using truncated kinesin-1 motors with a streptavidin-binding peptide tag that can attach to streptavidin-loaded, supported lipid bilayers. We found that the diffusing kinesin-1 motors propelled the microtubules in the presence of ATP. Notably, we found the gliding velocity of the microtubules to be strongly dependent on the number of motors and their diffusivity in the lipid bilayer. The microtubule gliding velocity increased with increasing motor density and membrane viscosity, reaching up to the stepping velocity of single motors. This finding is in contrast to conventional gliding motility assays where the density of surfaceimmobilized kinesin-1 motors does not influence the microtubule velocity over a wide range. We reason that the transport efficiency of membrane-anchored motors is reduced because of their slippage in the lipid bilayer, an effect that we directly observed using singlemolecule fluorescence microscopy. Our results illustrate the importance of motor-cargo coupling, which potentially provides cells with an additional means of regulating the efficiency of cargo transport. molecular motors | lipid bilayers | transport efficiency | motor-cargo coupling | streptavidin-binding peptide I ntracellular transport of membrane-bound vesicles and organelles is a process fundamental to many cellular functions including morphogenesis, signaling, and growth (1-4). Active cargo transport inside eukaryotic cells is mediated by ensembles of motor proteins, such as kinesins and dynein, walking on microtubule tracks (5), and myosins walking on actin filaments (6). Gaining mechanistic insight into the functioning of these motors inside the complex environment of cells is challenging. Several studies have thus used in vitro approaches to investigate transport mediated by groups of same or different motors attached to cargos such as silica beads (7), quantum dots (8), glass coverslips (9, 10), or DNA scaffolds (11,12). Although these approaches provide us with knowledge about the collective dynamics of multimotor transport, a key anomaly in these in vitro systems is the use of rather nonphysiological rigid cargo. Vesicular cargo transport by molecular motors requires their attachment to a fluid lipid bilayer either directly or via different adaptor molecules. The anchoring of motors in a diffusive lipid environment induces loose intermotor coupling along with the motors diffusing within the lipid bilayer, thereby increasing the flexibility of the system....

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Dynein Clusters into Lipid Microdomains on Phagosomes to Drive Rapid Transport toward Lysosomes

Cited by 195 publications

References 46 publications

Reggie-1 and reggie-2 (flotillins) participate in Rab11a-dependent cargo trafficking, spine synapse formation and LTP-related AMPA receptor (GluA1) surface exposure in mouse hippocampal neurons

Reggie-1 and reggie-2 (flotillins) participate in Rab11a-dependent cargo trafficking, spine synapse formation and LTP-related AMPA receptor (GluA1) surface exposure in mouse hippocampal neurons

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

Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity

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