The Experts below are selected from a list of 11757 Experts worldwide ranked by ideXlab platform
Nobutaka Hirokawa - One of the best experts on this subject based on the ideXlab platform.
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the molecular motor kif1a transports the trka neurotrophin receptor and is essential for sensory neuron survival and function
Neuron, 2016Co-Authors: Yosuke Tanaka, Shinsuke Niwa, Atena Farkhondeh, Ruyun Zhou, Ming Dong, Nobutaka Hirokawa, Li WangAbstract:Summary KIF1A is a major axonal transport motor protein, but its functional significance remains elusive. Here we show that KIF1A-haploinsufficient mice developed sensory neuropathy. We found progressive loss of TrkA(+) sensory neurons in Kif1a +/− dorsal root ganglia (DRGs). Moreover, axonal transport of TrkA was significantly disrupted in Kif1a +/− neurons. Live imaging and immunoprecipitation assays revealed that KIF1A bound to TrkA-containing vesicles through the adaptor GTP-Rab3, suggesting that TrkA is a cargo of the KIF1A motor. Physiological measurements revealed a weaker capsaicin response in Kif1a +/− DRG neurons. Moreover, these neurons were hyposensitive to nerve growth factor, which could explain the reduced neuronal survival and the functional deficiency of the pain receptor TRPV1. Because phosphatidylinositol 3-kinase (PI3K) signaling significantly rescued these phenotypes and also increased Kif1a mRNA, we propose that KIF1A is essential for the survival and function of sensory neurons because of the TrkA transport and its synergistic support of the NGF/TrkA/PI3K signaling pathway.
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Intracellular transport, molecular motors, KIFs and related diseases
BMC Genomics, 2014Co-Authors: Nobutaka HirokawaAbstract:The intracellular transport is fundamental for cellular functions, morphogenesis and survival in general including neurons composed of a very long axon and dendrites. We discovered most of the kinesin superfamily motor proteins, KIFs, 45 genes in mammals, elucidated their molecular structures and functional roles by molecular cell biology, molecular genetics, biophysics and structural biology and successfully disclosed the mechanism of intracellular transport fundamental for neuronal functions. In the axon and dendrites KIFs transport their cargos such as synaptic vesicle precursors (KIF1A/KIF1Bbeta), mitochondria (KIF1Balpha/KIF5s), plasma membrane proteins (KIF3/KIF5s), NMDA glutamate receptors (KIF17), AMPA receptors (KIF5s) and mRNA with large protein complex (KIF5s). KIFs mainly recognize and bind their cargoes through adaptor protein complex and release them via phosphorylation of KIFs or GTP hydrolyses of cargo G-protein. Furthermore, using molecular genetics we successfully uncovered that KIFs play significant roles for fundamental physiological phenomena in development and functions of nervous system and that deletion of KIFs causes certain diseases by clarifying followings: 1) KIF1A/KIF1B beta hetero mice serve as a model for neuropathy, 2) KIF3 determines left/right asymmetry by generating cilia and nodal flow in the node of early embryos, 3) KIF17 plays a fundamental role on learning and memory by not only transporting NMDA glutamate receptor in dendrites but also controlling transcription and translation of KIF17 and NMDA receptor mRNAs by enhancing phosphorylated CREB, 4) KIF1A is essential for hippocampal synaptogenesis and learning enhancement in an enriched environment, 5) KIF2A is fundamental for brain wiring by controlling unnecessary elongation of growth cones by depolymerizing microtubules, 6) KIF4 plays a crucial role in the activity-dependent survival of postmitotic neurons in brain development by regulating poly(ADP-ribose) polymerase-1 activity, 7) KIF26A is essential for enteric neuronal development by regulating GDNF-Ret signaling, 8) KIF3 suppresses tumorigenesis by transporting beta-catenin from Golgi to plasma membrane for serving as cell-cell adhesion molecules, inhibiting its accumulation in the nucleus and suppressing hyper proliferation of progenitor cells, 9)KIF5A is essential for GABAa receptor transport and KIF5A deletion causes epilepsy, 10)KIF19A is a microtubule depolymerizing KIF for ciliary length control and its deletion causes female infertility and hydrocephalus based on affected fluid flows, 11)KIF13A transports serotonin receptors to plasma membranes and its deletion causes elevated–anxiety phenotypes. Thus, KIFs play significant roles not only at cellular level, but also in brain function and development. Further, their malfunctions cause diseases such as neuropathy, epilepsy, dementia, elevated anxiety, tumor, megacolon and hydrocephalus.
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Regulation of NMDA Receptor Transport: A KIF17–Cargo Binding/Releasing Underlies Synaptic Plasticity and Memory In Vivo
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2012Co-Authors: Xiling Yin, Yosuke Takei, Xue Feng, Nobutaka HirokawaAbstract:Regulation of NMDA receptor trafficking is crucial to modulate neuronal communication. Ca 2+ /calmodulin-dependent protein kinase phosphorylates the tail domain of KIF17, a member of the kinesin superfamily, to control NMDA receptor subunit 2B (GluN2B) transport by changing the KIF17–cargo interaction in vitro . However, the mechanisms of regulation of GluN2B transport in vivo and its physiological significance are unknown. We generated transgenic mice carrying wild-type KIF17 ( TgS ), or KIF17 with S1029A ( TgA ) or S1029D ( TgD ) phosphomimic mutations in KIF17 −/− background. TgA/KIF17 −/− and TgD/KIF17 −/− mice exhibited reductions in synaptic NMDA receptors because of their inability to load/unload GluN2B onto/from KIF17, leading to impaired neuronal plasticity, CREB activation, and spatial memory. Expression of GFP-KIF17 in TgS/KIF17 −/− mouse neurons rescued the synaptic and behavioral defects of KIF17 −/− mice. These results suggest that phosphorylation-based regulation of NMDA receptor transport is critical for learning and memory in vivo .
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regulation of nmda receptor transport a KIF17 cargo binding releasing underlies synaptic plasticity and memory in vivo
The Journal of Neuroscience, 2012Co-Authors: Xiling Yin, Yosuke Takei, Nobutaka Hirokawa, Xue FengAbstract:Regulation of NMDA receptor trafficking is crucial to modulate neuronal communication. Ca 2+ /calmodulin-dependent protein kinase phosphorylates the tail domain of KIF17, a member of the kinesin superfamily, to control NMDA receptor subunit 2B (GluN2B) transport by changing the KIF17–cargo interaction in vitro . However, the mechanisms of regulation of GluN2B transport in vivo and its physiological significance are unknown. We generated transgenic mice carrying wild-type KIF17 ( TgS ), or KIF17 with S1029A ( TgA ) or S1029D ( TgD ) phosphomimic mutations in KIF17 −/− background. TgA/KIF17 −/− and TgD/KIF17 −/− mice exhibited reductions in synaptic NMDA receptors because of their inability to load/unload GluN2B onto/from KIF17, leading to impaired neuronal plasticity, CREB activation, and spatial memory. Expression of GFP-KIF17 in TgS/KIF17 −/− mouse neurons rescued the synaptic and behavioral defects of KIF17 −/− mice. These results suggest that phosphorylation-based regulation of NMDA receptor transport is critical for learning and memory in vivo .
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Motor protein KIF1A is essential for hippocampal synaptogenesis and learning enhancement in an enriched environment.
Neuron, 2012Co-Authors: Makoto Kondo, Yosuke Takei, Nobutaka HirokawaAbstract:Environmental enrichment causes a variety of effects on brain structure and function. Brain-derived neurotrophic factor (BDNF) plays an important role in enrichment-induced neuronal changes; however, the precise mechanism underlying these effects remains uncertain. In this study, a specific upregulation of kinesin superfamily motor protein 1A (KIF1A) was observed in the hippocampi of mice kept in an enriched environment and, in hippocampal neurons in vitro, BDNF increased the levels of KIF1A and of KIF1A-mediated cargo transport. Analysis of Bdnf(+/-) and Kif1a(+/-) mice revealed that a lack of KIF1A upregulation resulted in a loss of enrichment-induced hippocampal synaptogenesis and learning enhancement. Meanwhile, KIF1A overexpression promoted synaptogenesis via the formation of presynaptic boutons. These findings demonstrate that KIF1A is indispensable for BDNF-mediated hippocampal synaptogenesis and learning enhancement induced by enrichment. This is a new molecular motor-mediated presynaptic mechanism underlying experience-dependent neuroplasticity.
Hong Cao - One of the best experts on this subject based on the ideXlab platform.
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Role of spinal MCP-1-ERK-KIF17/NR2B signaling pathway in maintenance of type 2 diabetic neuropathic pain in rats
Chinese Journal of Anesthesiology, 2015Co-Authors: Jiayi Zhao, Hong CaoAbstract:Objective To explore the role of spinal monocyte chemoattractant protein-1 (MCP-1) -extracellular signal-regulated protein kinase (ERK) -kinesin superfamily motor protein 17 (KIF17) /N-methyl-D-aspartate receptor subunit 2B (NR2B) signaling pathway in the maintenance of type 2 diabetic neuropathic pain (DNP) in rats. Methods Type 2 diabetes mellitus was induced by a high-fat and high-sucrose diet and intraperitoneal streptozotocin (STZ) 35 mg/kg, and confirmed by fasting blood glucose level≥16.7 mmol/L 3 days later in male Sprague-Dawley rats aged 6 weeks.Type 2 DNP was confirmed when the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawl latency (TWL) measured on day 14 after STZ administration decreased to < 80% of the baseline value.The rats with type 2 DNP were randomly divided into 4 groups (n=36 each) using a random number table: type 2 DNP group (group DNP) , type 2 DNP + MCP-1 neutralizing antibody group (group DM) , type 2 DNP + ERK inhibitor group (group DE) and type 2 DNP + dimethyl sulfoxide group (group DD) . In DM, DE and DD groups, 0.1 ng/μl MCP-1 neutralizing antibody 10 μl, 0.5 μg/μl U0126 10 μl and 5 % dimethyl sulfoxide 10 μl were injected intrathecally, respectively, once a day for 14 consecutive days starting from 14 days after administration of STZ.Another 36 normal rats fed a common forage diet were adopted as control group (group C) . MWT and TWL were measured before STZ injection and at 1, 3, 7 and 14 days after STZ injection (T0-4) . Nine rats were sacrificed after measurement of pain thresholds at T1-4, and the lumbar segments (L4-6) of the spinal cord were removed for determination of the expression of phosphorylated ERK (p-ERK) , KIF17 and phosphorylated NR2B (p-NR2B) by Western blot. Results Compared with group C, the MWT was significantly decreased, the TWL was shortened, and the expression of p-ERK, KIF17 and p-NR2B was up-regulated at T1-4 in DNP, DM, DE and DD groups.Compared with group DNP, the MWT at T3-4 in group DM and at T2-4 in group DE was significantly increased, the TWL at T3-4 in group DM and at T2-4 in group DE was prolonged, and the expression of p-ERK, KIF17 and p-NR2B was down-regulated at T2-4 in DM and DE groups, and no significant changes were found in the parameters mentioned above in group DD. Conclusion Spinal MCP-1-ERK-KIF17/NR2B signaling pathway is involved in the maintenance of type 2 DNP in rats. Key words: Diabetes mellitus, type 2; Neuropathic pain; Chemokine CCL2; Extracellular Signal-Regulated MAP Kinases; Receptors, N-Methyl-D-Aspartate; Kinesin; Spinal cord
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role of spinal mcp 1 erk KIF17 nr2b signaling pathway in maintenance of type 2 diabetic neuropathic pain in rats
Chinese Journal of Anesthesiology, 2015Co-Authors: Jiayi Zhao, Hong CaoAbstract:Objective To explore the role of spinal monocyte chemoattractant protein-1 (MCP-1) -extracellular signal-regulated protein kinase (ERK) -kinesin superfamily motor protein 17 (KIF17) /N-methyl-D-aspartate receptor subunit 2B (NR2B) signaling pathway in the maintenance of type 2 diabetic neuropathic pain (DNP) in rats. Methods Type 2 diabetes mellitus was induced by a high-fat and high-sucrose diet and intraperitoneal streptozotocin (STZ) 35 mg/kg, and confirmed by fasting blood glucose level≥16.7 mmol/L 3 days later in male Sprague-Dawley rats aged 6 weeks.Type 2 DNP was confirmed when the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawl latency (TWL) measured on day 14 after STZ administration decreased to < 80% of the baseline value.The rats with type 2 DNP were randomly divided into 4 groups (n=36 each) using a random number table: type 2 DNP group (group DNP) , type 2 DNP + MCP-1 neutralizing antibody group (group DM) , type 2 DNP + ERK inhibitor group (group DE) and type 2 DNP + dimethyl sulfoxide group (group DD) . In DM, DE and DD groups, 0.1 ng/μl MCP-1 neutralizing antibody 10 μl, 0.5 μg/μl U0126 10 μl and 5 % dimethyl sulfoxide 10 μl were injected intrathecally, respectively, once a day for 14 consecutive days starting from 14 days after administration of STZ.Another 36 normal rats fed a common forage diet were adopted as control group (group C) . MWT and TWL were measured before STZ injection and at 1, 3, 7 and 14 days after STZ injection (T0-4) . Nine rats were sacrificed after measurement of pain thresholds at T1-4, and the lumbar segments (L4-6) of the spinal cord were removed for determination of the expression of phosphorylated ERK (p-ERK) , KIF17 and phosphorylated NR2B (p-NR2B) by Western blot. Results Compared with group C, the MWT was significantly decreased, the TWL was shortened, and the expression of p-ERK, KIF17 and p-NR2B was up-regulated at T1-4 in DNP, DM, DE and DD groups.Compared with group DNP, the MWT at T3-4 in group DM and at T2-4 in group DE was significantly increased, the TWL at T3-4 in group DM and at T2-4 in group DE was prolonged, and the expression of p-ERK, KIF17 and p-NR2B was down-regulated at T2-4 in DM and DE groups, and no significant changes were found in the parameters mentioned above in group DD. Conclusion Spinal MCP-1-ERK-KIF17/NR2B signaling pathway is involved in the maintenance of type 2 DNP in rats. Key words: Diabetes mellitus, type 2; Neuropathic pain; Chemokine CCL2; Extracellular Signal-Regulated MAP Kinases; Receptors, N-Methyl-D-Aspartate; Kinesin; Spinal cord
Joseph C Besharse - One of the best experts on this subject based on the ideXlab platform.
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KIF17 phosphorylation regulates photoreceptor outer segment turnover
BMC Cell Biology, 2018Co-Authors: Tylor R Lewis, Sean R Kundinger, Christine Insinna, Brian A Link, Joseph C BesharseAbstract:KIF17, a kinesin-2 motor that functions in intraflagellar transport, can regulate the onset of photoreceptor outer segment development. However, the function of KIF17 in a mature photoreceptor remains unclear. Additionally, the ciliary localization of KIF17 is regulated by a C-terminal consensus sequence (KRKK) that is immediately adjacent to a conserved residue (mouse S1029/zebrafish S815) previously shown to be phosphorylated by CaMKII. Yet, whether this phosphorylation can regulate the localization, and thus function, of KIF17 in ciliary photoreceptors remains unknown. Using transgenic expression in zebrafish photoreceptors, we show that phospho-mimetic KIF17 has enhanced localization along the cone outer segment. Importantly, expression of phospho-mimetic KIF17 is associated with greatly enhanced turnover of the photoreceptor outer segment through disc shedding in a cell-autonomous manner, while genetic mutants of KIF17 in zebrafish and mice have diminished disc shedding. Lastly, cone expression of constitutively active tCaMKII leads to a KIF17-dependent increase in disc shedding. Taken together, our data support a model in which phosphorylation of KIF17 promotes its photoreceptor outer segment localization and disc shedding, a process essential for photoreceptor maintenance and homeostasis. While disc shedding has been predominantly studied in the context of the mechanisms underlying phagocytosis of outer segments by the retinal pigment epithelium, this work implicates photoreceptor-derived signaling in the underlying mechanisms of disc shedding.
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KIF17 phosphorylation regulates its ciliary localization and photoreceptor outer segment turnover
bioRxiv, 2018Co-Authors: Tylor R Lewis, Sean R Kundinger, Christine Insinna, Brian A Link, Joseph C BesharseAbstract:Background: KIF17, a kinesin-2 motor that functions in intraflagellar transport, can regulate the onset of photoreceptor outer segment development. However, the function of KIF17 in a mature photoreceptor remains unclear. Additionally, the ciliary localization of KIF17 is regulated by a C-terminal consensus sequence (KRKK) that is immediately adjacent to a conserved residue (mouse S1029/zebrafish S815) previously shown to be phosphorylated by CaMKII. Yet, whether this phosphorylation can regulate the localization, and thus function, of KIF17 in ciliary photoreceptors remains unknown. Results: Using transgenic expression in both mammalian cells and zebrafish photoreceptors, we show that phospho-mimetic KIF17 has enhanced localization to cilia. Importantly, expression of phospho-mimetic KIF17 is associated with greatly enhanced turnover of the photoreceptor outer segment through disc shedding in a cell-autonomous manner, while genetic mutants of KIF17 in zebrafish and mice have diminished disc shedding. Lastly, cone expression of constitutively active tCaMKII leads to a KIF17-dependent increase in disc shedding. Conclusions: Taken together, our data support a model in which phosphorylation of KIF17 promotes its ciliary localization. In cone photoreceptor outer segments, this promotes disc shedding, a process essential for photoreceptor maintenance and homeostasis. While disc shedding has been predominantly studied in the context of the mechanisms underlying phagocytosis of outer segments by the retinal pigment epithelium, this work implicates photoreceptor-derived signaling in the underlying mechanisms of disc shedding.
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Additional file 1: of KIF17 phosphorylation regulates photoreceptor outer segment turnover
2018Co-Authors: Tylor Lewis, Sean R Kundinger, Christine Insinna, Brian Link, Joseph C BesharseAbstract:Figure S1. Phospho-mutations of S1029 regulate ciliary localization of KIF17. Figure S2. Phospho-mutations of S1029 mildly regulate nuclear localization of KIF17. Figure S3. Phospho-mutations of S815 regulate photoreceptor OS localization of KIF17. Figure S4. Mouse and zebrafish KIF17 are expressed rhythmically. Figure S5. Transient, episomal expression of phospho-mimetic KIF17(S815D) increases disc shedding. Figure S6. Opsin expression is largely unaffected by transgene expression. Figure S7. Rhodopsin immunogold labeling of phagosomes. Figure S8. Cone transducin-Îą immunogold labeling of phagosomes. Figure S9. Spline interpolation of zebrafish disc shedding data. Table S1. Spline interpolation of zebrafish disc shedding. Table S2. Spline interpolation of mouse disc shedding. (DOCX 5111 kb
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cos2 kif7 and osm 3 KIF17 regulate onset of outer segment development in zebrafish photoreceptors through distinct mechanisms
Developmental Biology, 2017Co-Authors: Tylor R Lewis, Sean R Kundinger, Brian A Link, Amira L Pavlovich, Jonathan R. Bostrom, Joseph C BesharseAbstract:Zebrafish morphants of osm-3/KIF17, a kinesin-2 family member and intraflagellar transport motor, have photoreceptor outer segments that are dramatically reduced in number and size. However, two genetic mutant lines, osm-3/KIF17sa0119 and osm-3/KIF17sa18340, reportedly lack any observable morphological outer segment defects. In this work, we use TALENs to generate an independent allele, osm-3/KIF17mw405, and show that both osm-3/KIF17sa0119 and osm-3/KIF17mw405 have an outer segment developmental delay in both size and density that is fully recovered by 6 days post-fertilization. Additionally, we use CRISPRs to generate cos2/kif7mw406, a mutation in the kinesin-4 family member cos2/kif7 that has been implicated in controlling ciliary architecture and Hedgehog signaling to test whether it may be functioning redundantly with osm-3/KIF17. We show that cos2/kif7mw406 has an outer segment developmental delay similar to the osm-3/KIF17 mutants. Using a three-dimensional mathematical model of outer segments, we show that while cos2/kif7mw406 and osm-3/KIF17mw405 outer segments are smaller throughout the first 6 days of development, the volumetric rates of outer segment morphogenesis are not different among wild-type, cos2/kif7mw406, and osm-3/KIF17mw405 after 60hpf. Instead, our model suggests that cos2/kif7mw406 and osm-3/KIF17mw405 impact outer segment morphogenesis through upstream events that that are different for each motor. In the case of cos2/kif7mw406 mutants, we show that early defects in Hedgehog signaling lead to a general, non-photoreceptor-specific delay of retinal neurogenesis, which in turn causes the secondary phenotype of delayed outer segment morphogenesis. In contrast, the osm-3/KIF17mw405 outer segment morphogenesis delays are linked specifically to initial disc morphogenesis of photoreceptors rather than an upstream event. Further, we show that osm-3/KIF17 mutant mice also exhibit a similarly delayed outer segment development, suggesting a role for osm-3/KIF17 in early outer segment development that is conserved across species. In conclusion, we show that both osm-3/KIF17 and cos2/kif7 have comparable outer segment developmental delays, although through independent mechanisms.
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Cos2/Kif7 and Osm-3/KIF17 regulate onset of outer segment development in zebrafish photoreceptors through distinct mechanisms.
Developmental biology, 2017Co-Authors: Tylor R Lewis, Sean R Kundinger, Brian A Link, Amira L Pavlovich, Jonathan R. Bostrom, Joseph C BesharseAbstract:Zebrafish morphants of osm-3/KIF17, a kinesin-2 family member and intraflagellar transport motor, have photoreceptor outer segments that are dramatically reduced in number and size. However, two genetic mutant lines, osm-3/KIF17sa0119 and osm-3/KIF17sa18340, reportedly lack any observable morphological outer segment defects. In this work, we use TALENs to generate an independent allele, osm-3/KIF17mw405, and show that both osm-3/KIF17sa0119 and osm-3/KIF17mw405 have an outer segment developmental delay in both size and density that is fully recovered by 6 days post-fertilization. Additionally, we use CRISPRs to generate cos2/kif7mw406, a mutation in the kinesin-4 family member cos2/kif7 that has been implicated in controlling ciliary architecture and Hedgehog signaling to test whether it may be functioning redundantly with osm-3/KIF17. We show that cos2/kif7mw406 has an outer segment developmental delay similar to the osm-3/KIF17 mutants. Using a three-dimensional mathematical model of outer segments, we show that while cos2/kif7mw406 and osm-3/KIF17mw405 outer segments are smaller throughout the first 6 days of development, the volumetric rates of outer segment morphogenesis are not different among wild-type, cos2/kif7mw406, and osm-3/KIF17mw405 after 60hpf. Instead, our model suggests that cos2/kif7mw406 and osm-3/KIF17mw405 impact outer segment morphogenesis through upstream events that that are different for each motor. In the case of cos2/kif7mw406 mutants, we show that early defects in Hedgehog signaling lead to a general, non-photoreceptor-specific delay of retinal neurogenesis, which in turn causes the secondary phenotype of delayed outer segment morphogenesis. In contrast, the osm-3/KIF17mw405 outer segment morphogenesis delays are linked specifically to initial disc morphogenesis of photoreceptors rather than an upstream event. Further, we show that osm-3/KIF17 mutant mice also exhibit a similarly delayed outer segment development, suggesting a role for osm-3/KIF17 in early outer segment development that is conserved across species. In conclusion, we show that both osm-3/KIF17 and cos2/kif7 have comparable outer segment developmental delays, although through independent mechanisms.
Kristen J Verhey - One of the best experts on this subject based on the ideXlab platform.
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Polyglutamylation of tubulin's C-terminal tail controls pausing and motility of kinesin-3 family member KIF1A.
Journal of Biological Chemistry, 2019Co-Authors: Dominique V. Lessard, Kristen J Verhey, Oraya J. Zinder, Takashi Hotta, Ryoma Ohi, Christopher L. BergerAbstract:The kinesin-3 family member KIF1A plays a critical role in site-specific neuronal cargo delivery during axonal transport. KIF1A cargo is mislocalized in many neurodegenerative diseases, indicating that KIF1A's highly efficient, superprocessive motility along axonal microtubules needs to be tightly regulated. One potential regulatory mechanism may be through posttranslational modifications (PTMs) of axonal microtubules. These PTMs often occur on the C-terminal tails of the microtubule tracks, act as molecular "traffic signals" helping to direct kinesin motor cargo delivery, and include C-terminal tail polyglutamylation important for KIF1A cargo transport. KIF1A initially interacts with microtubule C-terminal tails through its K-loop, a positively charged surface loop of the KIF1A motor domain. However, the role of the K-loop in KIF1A motility and response to perturbations in C-terminal tail polyglutamylation is underexplored. Using single-molecule imaging, we present evidence that KIF1A pauses on different microtubule lattice structures, linking multiple processive segments together and contributing to KIF1A's characteristic superprocessive run length. Furthermore, modifications of the KIF1A K-loop or tubulin C-terminal tail polyglutamylation reduced KIF1A pausing and overall run length. These results suggest a new mechanism to regulate KIF1A motility via pauses mediated by K-loop/polyglutamylated C-terminal tail interactions, providing further insight into KIF1A's role in axonal transport.
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Regulation of KIF1A motility via polyglutamylation of tubulin C-terminal tails
2018Co-Authors: Dominique V. Lessard, Kristen J Verhey, Oraya J. Zinder, Takashi Hotta, Ryoma Ohi, Christopher L. BergerAbstract:Axonal transport is a highly regulated cellular process responsible for site-specific neuronal cargo delivery. This process is mediated in part by KIF1A, a member of the kinesin-3 family of molecular motors. It is imperative that KIF1A9s highly efficient, superprocessive motility along microtubules is tightly regulated as misregulation of KIF1A cargo delivery is observed in many neurodegenerative diseases. However, the regulatory mechanisms responsible for KIF1A9s motility, and subsequent proper spatiotemporal cargo delivery, are largely unknown. One potential regulatory mechanism of KIF1A motility is through the posttranslational modifications (PTMs) of axonal microtubules. These PTMs, often occurring on the C-terminal tails of the microtubule tracks, act as molecular "traffic signals" helping to direct kinesin motor cargo delivery. Occurring on neuronal microtubules, C-terminal tail polygutamylation is known to be important for KIF1A cargo transport. KIF1A9s initial interaction with microtubule C-terminal tails is facilitated by the K-loop, a positively charged surface loop of the KIF1A motor domain. However, the K-loop9s role in KIF1A motility and response to perturbations in C-terminal tail polyglutamylation is underexplored. Using single-molecule imaging, we present evidence of KIF1A9s previously unreported pausing behavior on multiple microtubule structures. Further analysis revealed that these pauses link multiple processive segments together, contributing to KIF1A9s characteristic superprocessive run length. We further demonstrate that KIF1A pausing is mediated by a K-loop/polyglutamylated C-terminal tail interaction and is a regulatory mechanism of KIF1A motility. In summary, we introduce a new mechanism of KIF1A motility regulation, providing further insight into KIF1A9s role in axonal transport.
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dimerization of mammalian kinesin 3 motors results in superprocessive motion
Proceedings of the National Academy of Sciences of the United States of America, 2014Co-Authors: Virupakshi Soppina, Stephen R Norris, Aslan S Dizaji, Matt Kortus, Sarah L Veatch, Michelle Peckham, Kristen J VerheyAbstract:The kinesin-3 family is one of the largest among the kinesin superfamily and its members play important roles in a wide range of cellular transport activities, yet the molecular mechanisms of kinesin-3 regulation and cargo transport are largely unknown. We performed a comprehensive analysis of mammalian kinesin-3 motors from three different subfamilies (KIF1, KIF13, and KIF16). Using Forster resonance energy transfer microscopy in live cells, we show for the first time to our knowledge that KIF16B motors undergo cargo-mediated dimerization. The molecular mechanisms that regulate the monomer-to-dimer transition center around the neck coil (NC) segment and its ability to undergo intramolecular interactions in the monomer state versus intermolecular interactions in the dimer state. Regulation of NC dimerization is unique to the kinesin-3 family and in the case of KIF13A and KIF13B requires the release of a proline-induced kink between the NC and subsequent coiled-coil 1 segments. We show that dimerization of kinesin-3 motors results in superprocessive motion, with average run lengths of ∼10 μm, and that this property is intrinsic to the dimeric kinesin-3 motor domain. This finding opens up studies on the mechanistic basis of motor processivity. Such high processivity has not been observed for any other motor protein and suggests that kinesin-3 motors are evolutionarily adapted to serve as the marathon runners of the cellular world.
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ciliary entry of the kinesin 2 motor KIF17 is regulated by importin beta2 and rangtp
Nature Cell Biology, 2010Co-Authors: John F Dishinger, Jennetta W Hammond, Yen Nhu Thi Truong, Jeffrey R Martens, Toby W Hurd, Paul M Jenkins, Ben Margolis, Kristen J VerheyAbstract:How factors are targeted to cilia remains largely unknown. A ciliary localization signal targets the KIF17 motor, important for intraflagellar transport, to cilia through importin-β2 and RanGTP. The biogenesis, maintenance and function of primary cilia are controlled through intraflagellar transport (IFT) driven by two kinesin-2 family members, the heterotrimeric KIF3A/KIF3B/KAP complex and the homodimeric KIF17 motor1,2. How these motors and their cargoes gain access to the ciliary compartment is poorly understood. Here, we identify a ciliary localization signal (CLS) in the KIF17 tail domain that is necessary and sufficient for ciliary targeting. Similarities between the CLS and classic nuclear localization signals (NLSs) suggest that similar mechanisms regulate nuclear and ciliary import. We hypothesize that ciliary targeting of KIF17 is regulated by a ciliary-cytoplasmic gradient of the small GTPase Ran, with high levels of GTP-bound Ran (RanGTP) in the cilium. Consistent with this, cytoplasmic expression of GTP-locked Ran(G19V) disrupts the gradient and abolishes ciliary entry of KIF17. Furthermore, KIF17 interacts with the nuclear import protein importin-β2 in a manner dependent on the CLS and inhibited by RanGTP. We propose that Ran has a global role in regulating cellular compartmentalization by controlling the shuttling of cytoplasmic proteins into nuclear and ciliary compartments.
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single molecule imaging reveals differences in microtubule track selection between kinesin motors
PLOS Biology, 2009Co-Authors: Dawen Cai, Dyke P Mcewen, Jeffery R Martens, Edgar Meyhofer, Kristen J VerheyAbstract:Cells generate diverse microtubule populations by polymerization of a common α/β-tubulin building block. How microtubule associated proteins translate microtubule heterogeneity into specific cellular functions is not clear. We evaluated the ability of kinesin motors involved in vesicle transport to read microtubule heterogeneity by using single molecule imaging in live cells. We show that individual Kinesin-1 motors move preferentially on a subset of microtubules in COS cells, identified as the stable microtubules marked by post-translational modifications. In contrast, individual Kinesin-2 (KIF17) and Kinesin-3 (KIF1A) motors do not select subsets of microtubules. Surprisingly, KIF17 and KIF1A motors that overtake the plus ends of growing microtubules do not fall off but rather track with the growing tip. Selection of microtubule tracks restricts Kinesin-1 transport of VSVG vesicles to stable microtubules in COS cells whereas KIF17 transport of Kv1.5 vesicles is not restricted to specific microtubules in HL-1 myocytes. These results indicate that kinesin families can be distinguished by their ability to recognize microtubule heterogeneity. Furthermore, this property enables kinesin motors to segregate membrane trafficking events between stable and dynamic microtubule populations.
Yosuke Takei - One of the best experts on this subject based on the ideXlab platform.
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Regulation of NMDA Receptor Transport: A KIF17–Cargo Binding/Releasing Underlies Synaptic Plasticity and Memory In Vivo
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2012Co-Authors: Xiling Yin, Yosuke Takei, Xue Feng, Nobutaka HirokawaAbstract:Regulation of NMDA receptor trafficking is crucial to modulate neuronal communication. Ca 2+ /calmodulin-dependent protein kinase phosphorylates the tail domain of KIF17, a member of the kinesin superfamily, to control NMDA receptor subunit 2B (GluN2B) transport by changing the KIF17–cargo interaction in vitro . However, the mechanisms of regulation of GluN2B transport in vivo and its physiological significance are unknown. We generated transgenic mice carrying wild-type KIF17 ( TgS ), or KIF17 with S1029A ( TgA ) or S1029D ( TgD ) phosphomimic mutations in KIF17 −/− background. TgA/KIF17 −/− and TgD/KIF17 −/− mice exhibited reductions in synaptic NMDA receptors because of their inability to load/unload GluN2B onto/from KIF17, leading to impaired neuronal plasticity, CREB activation, and spatial memory. Expression of GFP-KIF17 in TgS/KIF17 −/− mouse neurons rescued the synaptic and behavioral defects of KIF17 −/− mice. These results suggest that phosphorylation-based regulation of NMDA receptor transport is critical for learning and memory in vivo .
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regulation of nmda receptor transport a KIF17 cargo binding releasing underlies synaptic plasticity and memory in vivo
The Journal of Neuroscience, 2012Co-Authors: Xiling Yin, Yosuke Takei, Nobutaka Hirokawa, Xue FengAbstract:Regulation of NMDA receptor trafficking is crucial to modulate neuronal communication. Ca 2+ /calmodulin-dependent protein kinase phosphorylates the tail domain of KIF17, a member of the kinesin superfamily, to control NMDA receptor subunit 2B (GluN2B) transport by changing the KIF17–cargo interaction in vitro . However, the mechanisms of regulation of GluN2B transport in vivo and its physiological significance are unknown. We generated transgenic mice carrying wild-type KIF17 ( TgS ), or KIF17 with S1029A ( TgA ) or S1029D ( TgD ) phosphomimic mutations in KIF17 −/− background. TgA/KIF17 −/− and TgD/KIF17 −/− mice exhibited reductions in synaptic NMDA receptors because of their inability to load/unload GluN2B onto/from KIF17, leading to impaired neuronal plasticity, CREB activation, and spatial memory. Expression of GFP-KIF17 in TgS/KIF17 −/− mouse neurons rescued the synaptic and behavioral defects of KIF17 −/− mice. These results suggest that phosphorylation-based regulation of NMDA receptor transport is critical for learning and memory in vivo .
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Motor protein KIF1A is essential for hippocampal synaptogenesis and learning enhancement in an enriched environment.
Neuron, 2012Co-Authors: Makoto Kondo, Yosuke Takei, Nobutaka HirokawaAbstract:Environmental enrichment causes a variety of effects on brain structure and function. Brain-derived neurotrophic factor (BDNF) plays an important role in enrichment-induced neuronal changes; however, the precise mechanism underlying these effects remains uncertain. In this study, a specific upregulation of kinesin superfamily motor protein 1A (KIF1A) was observed in the hippocampi of mice kept in an enriched environment and, in hippocampal neurons in vitro, BDNF increased the levels of KIF1A and of KIF1A-mediated cargo transport. Analysis of Bdnf(+/-) and Kif1a(+/-) mice revealed that a lack of KIF1A upregulation resulted in a loss of enrichment-induced hippocampal synaptogenesis and learning enhancement. Meanwhile, KIF1A overexpression promoted synaptogenesis via the formation of presynaptic boutons. These findings demonstrate that KIF1A is indispensable for BDNF-mediated hippocampal synaptogenesis and learning enhancement induced by enrichment. This is a new molecular motor-mediated presynaptic mechanism underlying experience-dependent neuroplasticity.
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molecular motor KIF17 is fundamental for memory and learning via differential support of synaptic nr2a 2b levels
Neuron, 2011Co-Authors: Yosuke Takei, Mizuho A Kido, Nobutaka HirokawaAbstract:Summary Kinesin superfamily motor protein 17 (KIF17) is a candidate transporter of N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B). Disruption of the murine KIF17 gene inhibits NR2B transport, accompanied by decreased transcription of nr2b , resulting in a loss of synaptic NR2B. In KIF17 −/− hippocampal neurons, the NR2A level is also decreased because of accelerated ubiquitin-proteasome system-dependent degradation. Accordingly, NMDA receptor-mediated synaptic currents, early and late long-term potentiation, long-term depression, and CREB responses are attenuated in KIF17 −/− neurons, concomitant with a hippocampus-dependent memory impairment in knockout mice. In wild-type neurons, CREB is activated by synaptic inputs, which increase the levels of KIF17 and NR2B. Thus, KIF17 differentially maintains the levels of NR2A and NR2B, and, when synapses are stimulated, the NR2B/KIF17 complex is upregulated on demand through CREB activity. These KIF17-based mechanisms for maintaining NR2A/2B levels could underlie multiple phases of memory processes in vivo.
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Molecular motor KIF17 is fundamental for memory and learning via differential support of synaptic NR2A/2B levels.
Neuron, 2011Co-Authors: Xiling Yin, Yosuke Takei, Mizuho A Kido, Nobutaka HirokawaAbstract:Summary Kinesin superfamily motor protein 17 (KIF17) is a candidate transporter of N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B). Disruption of the murine KIF17 gene inhibits NR2B transport, accompanied by decreased transcription of nr2b , resulting in a loss of synaptic NR2B. In KIF17 −/− hippocampal neurons, the NR2A level is also decreased because of accelerated ubiquitin-proteasome system-dependent degradation. Accordingly, NMDA receptor-mediated synaptic currents, early and late long-term potentiation, long-term depression, and CREB responses are attenuated in KIF17 −/− neurons, concomitant with a hippocampus-dependent memory impairment in knockout mice. In wild-type neurons, CREB is activated by synaptic inputs, which increase the levels of KIF17 and NR2B. Thus, KIF17 differentially maintains the levels of NR2A and NR2B, and, when synapses are stimulated, the NR2B/KIF17 complex is upregulated on demand through CREB activity. These KIF17-based mechanisms for maintaining NR2A/2B levels could underlie multiple phases of memory processes in vivo.
