KIF13A mediates the activity-dependent transport of ESCRT-0 proteins in axons

AbstractTurnover of synaptic vesicle (SV) proteins is vital for the maintenance of healthy, functional synapses in neurons. Our previous work showed that the degradation of SV proteins is mediated by the endosomal sorting complex required for transport (ESCRT) pathway in an activity-dependent manner. Here, we characterize the axonal transport dynamics of ESCRT-0 proteins Hrs and STAM1, the first components of the ESCRT pathway critical for initiating SV protein degradation. Hrs- and STAM1-positive transport vesicles exhibit increased anterograde and bidirectional motility in response to neuronal firing, and frequent colocalization with SV pools. These ESCRT-0 vesicles are a subset of Rab5-positive structures in axons, likely representing pro-degradative early endosomes. Further, we identify kinesin motor protein KIF13A as essential for the activity-dependent transport of ESCRT-0 vesicles as well as the degradation of SV membrane proteins. Together, these data demonstrate a novel KIF13A-dependent mechanism for mobilizing axonal transport of ESCRT machinery to facilitate the degradation of SV proteins.

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Arrestin-2 Interacts with the Endosomal Sorting Complex Required for Transport Machinery to Modulate Endosomal Sorting of CXCR4

Molecular Biology of the Cell ◽

10.1091/mbc.e10-02-0169 ◽

2010 ◽

Vol 21 (14) ◽

pp. 2529-2541 ◽

Cited By ~ 67

Author(s):

Rohit Malik ◽

Adriano Marchese

Keyword(s):

Molecular Mechanisms ◽

Lysosomal Degradation ◽

Kinase Substrate ◽

Endosomal Sorting ◽

Adaptor Molecule ◽

Escrt Machinery ◽

Early Endosomes ◽

G Protein Coupled ◽

Hepatocyte Growth ◽

Dependent Mechanism

The chemokine receptor CXCR4, a G protein-coupled receptor, is targeted for lysosomal degradation via a ubiquitin-dependent mechanism that involves the endosomal sorting complex required for transport (ESCRT) machinery. We have reported recently that arrestin-2 also targets CXCR4 for lysosomal degradation; however, the molecular mechanisms by which this occurs remain poorly understood. Here, we show that arrestin-2 interacts with ESCRT-0, a protein complex that recognizes and sorts ubiquitinated cargo into the degradative pathway. Signal-transducing adaptor molecule (STAM)-1, but not related STAM-2, interacts directly with arrestin-2 and colocalizes with CXCR4 on early endosomal antigen 1-positive early endosomes. Depletion of STAM-1 by RNA interference and disruption of the arrestin-2/STAM-1 interaction accelerates agonist promoted degradation of CXCR4, suggesting that STAM-1 via its interaction with arrestin-2 negatively regulates CXCR4 endosomal sorting. Interestingly, disruption of this interaction blocks agonist promoted ubiquitination of hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) but not CXCR4 and STAM-1 ubiquitination. Our data suggest a mechanism whereby arrestin-2 via its interaction with STAM-1 modulates CXCR4 sorting by regulating the ubiquitination status of HRS.

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A syntaxin 10–SNARE complex distinguishes two distinct transport routes from endosomes to the trans-Golgi in human cells

The Journal of Cell Biology ◽

10.1083/jcb.200707136 ◽

2008 ◽

Vol 180 (1) ◽

pp. 159-172 ◽

Cited By ~ 114

Author(s):

Ian G. Ganley ◽

Eric Espinosa ◽

Suzanne R. Pfeffer

Keyword(s):

Human Cells ◽

Endocytic Pathway ◽

Snare Complex ◽

Rab Gtpases ◽

Protein Receptor ◽

Transport Vesicles ◽

Early Endosomes ◽

Target Membrane ◽

Guanosine Triphosphatase ◽

Dependent Transport

Mannose 6-phosphate receptors (MPRs) are transported from endosomes to the Golgi after delivering lysosomal enzymes to the endocytic pathway. This process requires Rab9 guanosine triphosphatase (GTPase) and the putative tether GCC185. We show in human cells that a soluble NSF attachment protein receptor (SNARE) complex comprised of syntaxin 10 (STX10), STX16, Vti1a, and VAMP3 is required for this MPR transport but not for the STX6-dependent transport of TGN46 or cholera toxin from early endosomes to the Golgi. Depletion of STX10 leads to MPR missorting and hypersecretion of hexosaminidase. Mouse and rat cells lack STX10 and, thus, must use a different target membrane SNARE for this process. GCC185 binds directly to STX16 and is competed by Rab6. These data support a model in which the GCC185 tether helps Rab9-bearing transport vesicles deliver their cargo to the trans-Golgi and suggest that Rab GTPases can regulate SNARE–tether interactions. Importantly, our data provide a clear molecular distinction between the transport of MPRs and TGN46 to the trans-Golgi.

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Arf6 regulates the cycling and the readily releasable pool of synaptic vesicles at hippocampal synapse

eLife ◽

10.7554/elife.10116 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 24

Author(s):

Erica Tagliatti ◽

Manuela Fadda ◽

Antonio Falace ◽

Fabio Benfenati ◽

Anna Fassio

Keyword(s):

Plasma Membrane ◽

Synaptic Vesicles ◽

Small Gtpase ◽

Endosomal Sorting ◽

Releasable Pool ◽

Readily Releasable Pool ◽

Early Endosomes ◽

Hippocampal Synapses ◽

Activity Dependent ◽

Adp Ribosylation Factor

Recycling of synaptic vesicles (SVs) is a fundamental step in the process of neurotransmission. Endocytosed SV can travel directly into the recycling pool or recycle through endosomes but little is known about the molecular actors regulating the switch between these SV recycling routes. ADP ribosylation factor 6 (Arf6) is a small GTPase known to participate in constitutive trafficking between plasma membrane and early endosomes. Here, we have morphologically and functionally investigated Arf6-silenced hippocampal synapses and found an activity dependent accumulation of synaptic endosome-like organelles and increased release-competent docked SVs. These features were phenocopied by pharmacological blockage of Arf6 activation. The data reveal an unexpected role for this small GTPase in reducing the size of the readily releasable pool of SVs and in channeling retrieved SVs toward direct recycling rather than endosomal sorting. We propose that Arf6 acts at the presynapse to define the fate of an endocytosed SV.

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Faculty Opinions recommendation of Kinesin-1/Hsc70-dependent mechanism of slow axonal transport and its relation to fast axonal transport.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1787956.1562054 ◽

2010 ◽

Author(s):

Orly Reiner

Keyword(s):

Axonal Transport ◽

Slow Axonal Transport ◽

Fast Axonal Transport ◽

Dependent Mechanism

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The Novel ALG-2 Target Protein CDIP1 Promotes Cell Death by Interacting with ESCRT-I and VAPA/B

International Journal of Molecular Sciences ◽

10.3390/ijms22031175 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1175

Author(s):

Ryuta Inukai ◽

Kanako Mori ◽

Keiko Kuwata ◽

Chihiro Suzuki ◽

Masatoshi Maki ◽

...

Keyword(s):

Cell Death ◽

Essential Gene ◽

Susceptibility Gene ◽

Target Protein ◽

Hek293 Cells ◽

Dependent Manner ◽

Gene Encoding ◽

Endosomal Sorting ◽

Cell Death Pathways ◽

Apoptotic Protein

Apoptosis-linked gene 2 (ALG-2, also known as PDCD6) is a member of the penta-EF-hand (PEF) family of Ca2+-binding proteins. The murine gene encoding ALG-2 was originally reported to be an essential gene for apoptosis. However, the role of ALG-2 in cell death pathways has remained elusive. In the present study, we found that cell death-inducing p53 target protein 1 (CDIP1), a pro-apoptotic protein, interacts with ALG-2 in a Ca2+-dependent manner. Co-immunoprecipitation analysis of GFP-fused CDIP1 (GFP-CDIP1) revealed that GFP-CDIP1 associates with tumor susceptibility gene 101 (TSG101), a known target of ALG-2 and a subunit of endosomal sorting complex required for transport-I (ESCRT-I). ESCRT-I is a heterotetrameric complex composed of TSG101, VPS28, VPS37 and MVB12/UBAP1. Of diverse ESCRT-I species originating from four VPS37 isoforms (A, B, C, and D), CDIP1 preferentially associates with ESCRT-I containing VPS37B or VPS37C in part through the adaptor function of ALG-2. Overexpression of GFP-CDIP1 in HEK293 cells caused caspase-3/7-mediated cell death. In addition, the cell death was enhanced by co-expression of ALG-2 and ESCRT-I, indicating that ALG-2 likely promotes CDIP1-induced cell death by promoting the association between CDIP1 and ESCRT-I. We also found that CDIP1 binds to vesicle-associated membrane protein-associated protein (VAP)A and VAPB through the two phenylalanines in an acidic tract (FFAT)-like motif in the C-terminal region of CDIP1, mutations of which resulted in reduction of CDIP1-induced cell death. Therefore, our findings suggest that different expression levels of ALG-2, ESCRT-I subunits, VAPA and VAPB may have an impact on sensitivity of anticancer drugs associated with CDIP1 expression.

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Extracellular Release of ILEI/FAM3C and Amyloid-β Is Associated with the Activation of Distinct Synapse Subpopulations

Journal of Alzheimer s Disease ◽

10.3233/jad-201174 ◽

2021 ◽

pp. 1-16

Author(s):

Masaki Nakano ◽

Yachiyo Mitsuishi ◽

Lei Liu ◽

Naoki Watanabe ◽

Emi Hibino ◽

...

Keyword(s):

Neuronal Activity ◽

Epithelial Mesenchymal Transition ◽

Surrogate Marker ◽

Amyloid Β ◽

Dependent Manner ◽

Mesenchymal Transition ◽

Extracellular Release ◽

Activity Dependent ◽

Aβ Production

Background: Brain amyloid-β (Aβ) peptide is released into the interstitial fluid (ISF) in a neuronal activity-dependent manner, and Aβ deposition in Alzheimer’s disease (AD) is linked to baseline neuronal activity. Although the intrinsic mechanism for Aβ generation remains to be elucidated, interleukin-like epithelial-mesenchymal transition inducer (ILEI) is a candidate for an endogenous Aβ suppressor. Objective: This study aimed to access the mechanism underlying ILEI secretion and its effect on Aβ production in the brain. Methods: ILEI and Aβ levels in the cerebral cortex were monitored using a newly developed ILEI-specific ELISA and in vivo microdialysis in mutant human Aβ precursor protein-knockin mice. ILEI levels in autopsied brains and cerebrospinal fluid (CSF) were measured using ELISA. Results: Extracellular release of ILEI and Aβ was dependent on neuronal activation and specifically on tetanus toxin-sensitive exocytosis of synaptic vesicles. However, simultaneous monitoring of extracellular ILEI and Aβ revealed that a spontaneous fluctuation of ILEI levels appeared to inversely mirror that of Aβ levels. Selective activation and inhibition of synaptic receptors differentially altered these levels. The evoked activation of AMPA-type receptors resulted in opposing changes to ILEI and Aβ levels. Brain ILEI levels were selectively decreased in AD. CSF ILEI concentration correlated with that of Aβ and were reduced in AD and mild cognitive impairment. Conclusion: ILEI and Aβ are released from distinct subpopulations of synaptic terminals in an activity-dependent manner, and ILEI negatively regulates Aβ production in specific synapse types. CSF ILEI might represent a surrogate marker for the accumulation of brain Aβ.

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Specific release of membrane-bound annexin II and cortical cytoskeletal elements by sequestration of membrane cholesterol.

Molecular Biology of the Cell ◽

10.1091/mbc.8.3.533 ◽

1997 ◽

Vol 8 (3) ◽

pp. 533-545 ◽

Cited By ~ 159

Author(s):

T Harder ◽

R Kellner ◽

R G Parton ◽

J Gruenberg

Keyword(s):

Actin Cytoskeleton ◽

Cytoskeletal Proteins ◽

Annexin Ii ◽

Dependent Manner ◽

Cortical Actin ◽

Independent Manner ◽

Low Concentrations ◽

Early Endosomes ◽

Associated Proteins ◽

Cytoskeletal Elements

Annexin II is an abundant protein which is present in the cytosol and on the cytoplasmic face of plasma membrane and early endosomes. It is generally believed that this association occurs via Ca(2+)-dependent binding to lipids, a mechanism typical for the annexin protein family. Although previous studies have shown that annexin II is involved in early endosome dynamics and organization, the precise biological role of the protein is unknown. In this study, we found that approximately 50% of the total cellular annexin was associated with membranes in a Ca(2+)-independent manner. This binding was extremely tight, since it resisted high salt and, to some extent, high pH treatments. We found, however, that membrane-associated annexin II could be quantitatively released by low concentrations of the cholesterol-sequestering agents filipin and digitonin. Both treatments released an identical and limited set of proteins but had no effects on other membrane-associated proteins. Among the released proteins, we identified, in addition to annexin II itself, the cortical cytoskeletal proteins alpha-actinin, ezrin and moesin, and membrane-associated actin. Our biochemical and immunological observations indicate that these proteins are part of a complex containing annexin II and that stability of the complex is sensitive to cholesterol sequestering agents. Since annexin II is tightly membrane-associated in a cholesterol-dependent manner, and since it seems to interact physically with elements of the cortical actin cytoskeleton, we propose that the protein serves as interface between membranes containing high amounts of cholesterol and the actin cytoskeleton.

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Retrograde Axonal Transport: a Major Transmission Route of Enterovirus 71 in Mice

Journal of Virology ◽

10.1128/jvi.00236-07 ◽

2007 ◽

Vol 81 (17) ◽

pp. 8996-9003 ◽

Cited By ~ 111

Author(s):

Che-Szu Chen ◽

Yi-Chuan Yao ◽

Shin-Chao Lin ◽

Yi-Ping Lee ◽

Ya-Fang Wang ◽

...

Keyword(s):

Axonal Transport ◽

Early Stage ◽

Clinical Signs ◽

Enterovirus 71 ◽

Severe Infection ◽

Dependent Manner ◽

Transmission Route ◽

Brain Infection ◽

Reverse Pattern ◽

Clinical Illness

ABSTRACT Inoculation of enterovirus 71 (EV71) by the oral (p.o.), intramuscular (i.m.), or intracranial route resulted in brain infection, flaccid paralysis, pulmonary dysfunction, and death of 7-day-old mice. The lag time of disease progression indicated that neuroinvasion from the inoculation sites was a prerequisite for the development of the clinical signs. Although EV71 p.o. inoculation led to a persistent viremia and a transient increase in blood-brain barrier permeability at the early stage of the infection, only low levels of virus, which led to neither severe infection nor clinical illness, could be detected in the brain, suggesting that hematogenous transport might not represent a major transmission route. In the spinal cord, following both p.o. and hind limb i.m. inoculation, the virus first appeared and increased rapidly in the lower segments, especially at the anterior horn areas, and then spread to the upper segments and brain in the presence of viremia. A reverse pattern, with the virus being first detected in the upper segment, was observed when the virus was i.m. inoculated in the forelimb. Colchicine, a fast axonal transport inhibitor, but not sciatic nerve transection reduced EV71 neuroinvasion in a dose-dependent manner, indicating a neuronal transmission of the virus.

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The Carbamate, Physostigmine does not Impair Axonal Transport in Rat Cortical Neurons

Neuroscience Insights ◽

10.1177/26331055211020289 ◽

2021 ◽

Vol 16 ◽

pp. 263310552110202

Author(s):

Sean X Naughton ◽

Wayne D Beck ◽

Zhe Wei ◽

Guangyu Wu ◽

Peter W Baas ◽

...

Keyword(s):

Axonal Transport ◽

Cortical Neurons ◽

Acetylcholinesterase Inhibitors ◽

Neurological Deficits ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Rat Cortical Neurons ◽

Toxicological Mechanism

Among the various chemicals that are commonly used as pesticides, organophosphates (OPs), and to a lesser extent, carbamates, are most frequently associated with adverse long-term neurological consequences. OPs and the carbamate, pyridostigmine, used as a prophylactic drug against potential nerve agent attacks, have also been implicated in Gulf War Illness (GWI), which is often characterized by chronic neurological symptoms. While most OP- and carbamate-based pesticides, and pyridostigmine are relatively potent acetylcholinesterase inhibitors (AChEIs), this toxicological mechanism is inadequate to explain their long-term health effects, especially when no signs of acute cholinergic toxicity are exhibited. Our previous work suggests that a potential mechanism of the long-term neurological deficits associated with OPs is impairment of axonal transport (AXT); however, we had not previously evaluated carbamates for this effect. Here we thus evaluated the carbamate, physostigmine (PHY), a highly potent AChEI, on AXT using an in vitro neuronal live imaging assay that we have previously found to be very sensitive to OP-related deficits in AXT. We first evaluated the OP, diisopropylfluorophosphate (DFP) (concentration range 0.001-10.0 µM) as a reference compound that we found previously to impair AXT and subsequently evaluated PHY (concentration range 0.01-100 nM). As expected, DFP impaired AXT in a concentration-dependent manner, replicating our previously published results. In contrast, none of the concentrations of PHY (including concentrations well above the threshold for impairing AChE) impaired AXT. These data suggest that the long-term neurological deficits associated with some carbamates are not likely due to acute impairments of AXT.

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