Recombinant Globular Tail Fragment of Myosin-V Blocks Vesicle Transport in Squid Nerve Cell Extracts

Brown, Jeremiah R.; Simonetta, K.R.; Sandberg, Leslie A.; Stafford, Phillip; Langford, George M.

doi:10.2307/1543343

Cited by 7 publications

(12 citation statements)

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“…3A) to knockdown endogenous Myo5a. To control for off-target effects, we also expressed Myo5aCBD, which acts as a dominant-negative form because it associates with TrkB-FL but lacks the motor domain (Brown et al, 2001). First, we used a live-cell ratiometric fluorescence-based endocytosis assay (Kotowski et al, 2011) to analyze TrkB-FL internalization in cultured hippocampal neurons (Fig.…”

Section: Myo5a Is Not Involved In Bdnf-dependent Trkb-fl Internalizationmentioning

confidence: 99%

Myosin5a mediates BDNF-induced postendocytic recycling of full-length TrkB and its translocation into dendritic spines

Sui

Huang

Wang

et al. 2015

Journal of Cell Science

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Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival, neurite outgrowth and synaptic plasticity by activating the receptor tropomyosin receptor kinase B (TrkB, also known as NTRK2). TrkB has been shown to undergo recycling after BDNF stimulation. We have previously reported that full-length TrkB (TrkB-FL) are recycled through a Rab11-dependent pathway upon BDNF stimuli, which is important for the translocation of TrkB-FL into dendritic spines and for the maintenance of prolonged BDNF downstream signaling during long-term potentiation (LTP). However, the identity of the motor protein that mediates the local transfer of recycled TrkB-FL back to the plasma membrane remains unclear. Here, we report that the F-actin-based motor protein myosin Va (Myo5a) mediates the postendocytic recycling of TrkB-FL. Blocking the interaction between Rab11 and Myo5a by use of a TAT-tagged peptide consisting of amino acids 55-66 of the Myo5a ExonE domain weakened the association between TrkB-FL and Myo5a and thus impaired TrkB-FL recycling and BDNF-induced TrkB-FL translocation into dendritic spines. Finally, inhibiting Myo5a-mediated TrkB-FL recycling led to a significant reduction in prolonged BDNF downstream signaling. Taken together, these results show that Myo5a mediates BDNF-dependent TrkB-FL recycling and contributes to BDNF-induced TrkB spine translocation and prolonged downstream signaling.

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Section: Myo5a Is Not Involved In Bdnf-dependent Trkb-fl Internalizationmentioning

confidence: 99%

Myosin5a mediates BDNF-induced postendocytic recycling of full-length TrkB and its translocation into dendritic spines

Sui

Huang

Wang

et al. 2015

Journal of Cell Science

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“…In addition, biochemical studies and immunofluorescence microscopy have been used to verify the localization of myosin‐V on vesicles (12,33–35). In vitro motility assays, in which vesicle movement on actin filaments is inhibited by myosin‐V specific probes (antibodies, recombinant protein fragments) provide evidence for a role in vesicle movement (33,36–39). Ultimately, the reconstitution of movement from biochemically purified components will be needed to determine the protein composition of active motor complexes on vesicles.…”

Section: Vesicle Transport By Myosin‐vmentioning

confidence: 99%

“…In MDCK cells, the tail fragment of myosin‐Vb retards trafficking of vesicles and, in HeLa cells, the tail fragment of myosin‐Vc disrupts the distribution of endosomal vesicles containing transferrin receptors (12,31). In extruded axoplasm of the squid giant axon, a bacterially expressed mouse globular tail fragment blocks vesicle transport on actin filaments (38,39).…”

Section: Recruitment Of Myosin‐v To Vesicles By the Rab/rabphilin Commentioning

confidence: 99%

“…An exciting recent discovery is the finding that the cargo‐binding domain of myosin‐V interacts with kinesin, a microtubule‐based motor (Figure 2) (38,39,56,57). The interaction of myosin‐V with kinesin has been established through several different approaches: yeast 2‐hybrid assay, coimmunoprecipitation, coaffinity isolation, and copurification with myosin‐V.…”

Section: Myosin‐v/kinesin Interactionsmentioning

confidence: 99%

“…The distal/globular tail domain of myosin‐V binds to the rod‐tail domain of kinesin to form a ‘heteromotor’ complex. The members of the kinesin super‐family thus far shown to act as binding partners of myosin‐V include conventional or ubiquitous kinesin and Smy1p (38,39,56,57). Evidence that myosin‐V and kinesin interact on the membrane surface has not yet been demonstrated.…”

Section: Myosin‐v/kinesin Interactionsmentioning

confidence: 99%

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Myosin‐V, a Versatile Motor for Short‐Range Vesicle Transport

Langford¹

2002

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Myosin-V is a versatile motor involved in short-range transport of vesicles in the actin-rich cortex of the cell. It binds to several different kinds of vesicles, and the mechanism by which it interacts with the vesicle surface is being unraveled, primarily in melanocytes. Members of the Rab family of G-proteins are required for the recruitment of myosin-V to vesicles. Rab27a and its rabphilin-like effector protein, Melanophilin, recruit myosin-Va to melanosomes and appear to serve as the membrane receptor. Myosin-V is also involved in fast axonal/dendritic transport and, interestingly, it forms a complex with kinesin, a microtubule-based motor. This kinesin/myosin-V heteromotor complex allows long-range movement of vesicles within axons and dendrites on microtubules and short-range movement in the dendritic spines and axon terminals on actin filaments. The direct interaction of motors from both filament systems may represent the mechanism by which the transition of vesicles from microtubules to actin filaments is regulated. The actin-based molecular motors constitute a large superfamily of motor proteins, collectively called myosin (1). Myosin-V, one of 18 known classes of myosin motors, is involved in fast axonal/dendritic transport, the anterograde and retrograde movement of vesicular cargo in neurons. Our understanding of the functions of myosin-V in axonal transport has grown rapidly over the past decade. It has been established that the principal cargo transported by myosin-V in neurons and other cell types are vesicles/organelles [for reviews see (2-4)]. Vesicles are abundant in cells and motor-mediated transport is required for proper distribution. Myosin-V is absolutely essential for transport of vesicles in actin-rich cortical regions of neurons, i.e. dendritic spines and axon termini. Moreover, vesicle-associated myosins, like myosin-V, may do more than serve as 'mechanical feet' transporting vesicles from one location to another along actin tracks. They may interact with membrane-associated F-actin to 'stitch' mem-859 branes together during membrane fusion (5) and to 'sort' proteins from the fluid phase into lipid rafts during the assembly of membrane protein complexes (6). The secret to the versatility of molecular motors lies in the diversity of proteins that interact with them and that function to recruit them to the appropriate membrane compartments. This review will cover recent studies of myosin-V and will focus on its cargobinding domain.Based on phylogenetic analysis of the myosin super-family, myosin-V evolved after myosins-II and I, the two most ancient members of the myosin super-family (7-9). Myosin-V is broadly distributed in the fungi and animal phyla but is missing from plants. However, myosin-XI, a class of myosin closely related to myosin-V, is found in plants (10,11). Remarkably, myosins-V and XI show high sequence homology, share structural features (10), and exhibit similar functions (7). This suggests that myosin-XI and V represent two distinct members of a broadly defined V/XI c...

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Short‐range axonal/dendritic transport by myosin‐V: A model for vesicle delivery to the synapse

2003

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Myosin-V is a versatile motor involved in short-range axonal/dendritic transport of vesicles in the actin-rich cortex and synaptic regions of nerve cells. It binds to several different kinds of neuronal vesicles by its globular tail domain but the mechanism by which it is recruited to these vesicles is not known. In this study, we used an in vitro motility assay derived from axoplasm of the squid giant axon to study the effects of the globular tail domain on the transport of neuronal vesicles. We found that the globular tail fragment of myosin-V inhibited actin-based vesicle transport by displacing native myosin-V and binding to vesicles. The globular tail domain pulled down kinesin, a known binding partner of myosin-V, in affinity isolation experiments. These data confirmed earlier evidence that kinesin and myosin-V interact to form a hetero-motor complex. The formation of a kinesin/myosin-V hetero-motor complex on vesicles is thought to facilitate the coordination of long-range movement on microtubules and short-range movement on actin filaments. The direct interaction of motors from both filament systems may represent the mechanism by which the transition of vesicles from microtubules to actin filaments is regulated. These results are the first demonstration that the recombinant tail of myosin-V inhibits vesicle transport in an in vitro motility assay. Future experiments are designed to determine the functional significance of the interaction between myosin-V and kinesin and to identify other proteins that bind to the globular tail domain of myosin-V.

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Recombinant Globular Tail Fragment of Myosin-V Blocks Vesicle Transport in Squid Nerve Cell Extracts

Cited by 7 publications

References 2 publications

Myosin5a mediates BDNF-induced postendocytic recycling of full-length TrkB and its translocation into dendritic spines

Myosin5a mediates BDNF-induced postendocytic recycling of full-length TrkB and its translocation into dendritic spines

Myosin‐V, a Versatile Motor for Short‐Range Vesicle Transport

Short‐range axonal/dendritic transport by myosin‐V: A model for vesicle delivery to the synapse

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