Myosin-Va and Dynamic Actin Oppose Microtubules to Drive Long-Range Organelle Transport

Evans, Richard D.; Robinson, Christopher L.; Briggs, Deborah A.; Tooth, David; Ramalho, José S.; Cantero, Marta; Montoliu, Lluı́s; Patel, Shyamal; Sviderskaya, Elena V.; Hume, Alistair N.

doi:10.1016/j.cub.2014.06.019

Cited by 60 publications

(72 citation statements)

References 36 publications

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“…It is reported for melanocytes that both kinesin and myosin Va are involved in the transportation and secretion of melanosomes 50–52 . Identification of myosin motors responsible for Col-1 secretion requires further study.…”

Section: Discussionmentioning

confidence: 99%

KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis

Kamata

Tsukasaki

Sakai

et al. 2017

Sci Rep

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Fibrosis involves the production of extracellular matrix proteins in tissues and is often preceded by injury or trauma. In pleural fibrosis excess collagen deposition results in pleural thickening, increased stiffness and impaired lung function. Myofibroblasts are responsible for increased collagen deposition, however the molecular mechanism of transportation of procollagen containing vesicles for secretion is unknown. Here, we studied the role of kinesin on collagen-1 (Col-1) containing vesicle transportation in human pleural mesothelial cells (HPMCs). Among a number of cargo transporting kinesins, KIF5A was notably upregulated during TGF-β induced mesothelial-mesenchymal transition (MesoMT). Using superresolution structured illumination microscopy and the DUO-Link technique, we found that KIF5A colocalized with Col-1 containing vesicles. KIF5A knock-down significantly reduced Col-1 secretion and attenuated TGF-β induced increment in Col-1 localization at cell peripheries. Live cell imaging revealed that GFP-KIF5A and mCherry-Col-1 containing vesicles moved together. Kymography showed that these molecules continuously move with a mean velocity of 0.56 μm/sec, suggesting that the movement is directional but not diffusion limited process. Moreover, KIF5A was notably upregulated along with Col-1 and α-smooth muscle actin in pleural thickening in the carbon-black bleomycin mouse model. These results support our hypothesis that KIF5A is responsible for collagen transportation and secretion from HPMCs.

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Section: Discussionmentioning

confidence: 99%

KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis

Kamata

Tsukasaki

Sakai

et al. 2017

Sci Rep

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“…MyoVa function, on the other hand, was indispensable for the peripheral distribution of melanosomes on the actin network (5-7, 28, 29). Depolymerization of the actin network or the absence of MyoVa resulted in a microtubule-dependent clustering of melanosomes in the cell center (28,30,31). However, the signaling pathway(s) that regulate the directionality of the melanosome transport on the microtubule and actin networks have not yet been delineated.…”

Section: Significancementioning

confidence: 99%

Myosin Va’s adaptor protein melanophilin enforces track selection on the microtubule and actin networks in vitro

Oberhofer

Spieler

Rosenfeld

et al. 2017

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

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Pigment organelles, or melanosomes, are transported by kinesin, dynein, and myosin motors. As such, melanosome transport is an excellent model system to study the functional relationship between the microtubule-and actin-based transport systems. In mammalian melanocytes, it is well known that the Rab27a/melanophilin/myosin Va complex mediates actin-based transport in vivo. However, pathways that regulate the overall directionality of melanosomes on the actin/microtubule networks have not yet been delineated. Here, we investigated the role of PKA-dependent phosphorylation on the activity of the actin-based Rab27a/melanophilin/myosin Va transport complex in vitro. We found that melanophilin, specifically its C-terminal actin-binding domain (ABD), is a target of PKA. Notably, in vitro phosphorylation of the ABD closely recapitulated the previously described in vivo phosphorylation pattern. Unexpectedly, we found that phosphorylation of the ABD affected neither the interaction of the complex with actin nor its movement along actin tracks. Surprisingly, the phosphorylation state of melanophilin was instead important for reversible association with microtubules in vitro. Dephosphorylated melanophilin preferred binding to microtubules even in the presence of actin, whereas phosphorylated melanophilin associated with actin. Indeed, when actin and microtubules were present simultaneously, melanophilin's phosphorylation state enforced track selection of the Rab27a/melanophilin/ myosin Va transport complex. Collectively, our results unmasked the regulatory dominance of the melanophilin adaptor protein over its associated motor and offer an unexpected mechanism by which filaments of the cytoskeletal network compete for the moving organelles to accomplish directional transport on the cytoskeleton in vivo.melanophilin | myosin Va | intracellular transport | transport regulation

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“…Evans et al. indicated that MyosinVa (MyoVa) and actin are another main drivers of melanosome dispersion in murine melanocytes …”

Section: Introductionmentioning

confidence: 99%

The absence of Rab27a accelerates the degradation of Melanophilin

Park

Lee

Myung

et al. 2019

Experimental Dermatology

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Melanophilin (Mlph) forms an interaction with Rab27a and the actin‐based motor protein MyosinVa (MyoVa) on mature melanosome membranes and the tripartite complex regulates melanosome transport in melanocytes. In this study, we found that Rab27a siRNA decreased Mlph and Rab27a protein levels, but Mlph mRNA levels were not changed. Other Rab27a siRNA sequences also showed the same results. When Rab27a siRNA was treated with melan‐a melanocytes, Rab27a protein was decreased within 6 hours and Mlph protein was decreased within 24 hours. To determine whether the absence of Rab27a promotes Mlph degradation, we inhibited protein degradation by treatment with proteasome (MG132) and lysosomal enzyme (E64D and Pepstatin A) inhibitors in melan‐a melanocytes. MG132 inhibited the degradation of Mlph, but E64D and Pepstatin A had no effect on Mlph. The absence of Rab27a enhanced ubiquitination of Mlph and induced proteasomal degradation. From these results, we concluded that Mlph interaction with Rab27a is important for Mlph stability and melanosome transport.

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Myosin-Va and Dynamic Actin Oppose Microtubules to Drive Long-Range Organelle Transport

Cited by 60 publications

References 36 publications

KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis

KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis

Myosin Va’s adaptor protein melanophilin enforces track selection on the microtubule and actin networks in vitro

The absence of Rab27a accelerates the degradation of Melanophilin

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