Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Yang, Rui; Bentley, Marvin; Huang, Chung Fang; Banker, Gary

doi:10.1016/bs.mcb.2015.06.021

Cited by 11 publications

(15 citation statements)

References 40 publications

(70 reference statements)

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“…Figure A shows example kymographs illustrating the movements of two different kinesins (KIF5B and KIF13A), each co‐expressed with the axonal plasma membrane protein neuron‐glia cell adhesion molecule (NgCAM) . Previous work and the behavior of KIF5 tails shown here (Figure ) support the hypothesis that members of the Kinesin‐1 family mediate the transport of axonally polarized proteins. However, this kinesin‐cargo interaction has never been shown directly.…”

Section: Resultssupporting

confidence: 62%

“…Much is known about how kinesin motor domains behave in vitro and in living cells, but relatively little is known about the behavior of vesicle‐bound kinesins. One surprising outcome from this study is that only six kinesins, all members of the Kinesin‐1 and Kinesin‐3 families, exhibited long‐range transport.…”

Section: Discussionmentioning

confidence: 99%

“…Other kinesins labeled distinct populations of somatodendritic organelles that seldom underwent long‐range movements, perhaps indicating a function for these kinesins beyond long‐range transport. Notably, the preference of vesicle‐attached kinesins for axons or dendrites often differed from the transport preference of their motor domains, as defined in motor‐domain expression assays . This observation suggests that the identity of the vesicle that a kinesin binds is a crucial determinant of that kinesin's transport parameters, an idea we refer to as “on‐vesicle regulation”.…”

Section: Introductionmentioning

confidence: 99%

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A novel strategy to visualize vesicle‐bound kinesins reveals the diversity of kinesin‐mediated transport

Yang

Bostick

Garbouchian

et al. 2019

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In mammals, 15 to 20 kinesins are thought to mediate vesicle transport. Little is known about the identity of vesicles moved by each kinesin or the functional significance of such diversity. To characterize the transport mediated by different kinesins, we developed a novel strategy to visualize vesicle‐bound kinesins in living cells. We applied this method to cultured neurons and systematically determined the localization and transport parameters of vesicles labeled by different members of the Kinesin‐1, ‐2, and ‐3 families. We observed vesicle labeling with nearly all kinesins. Only six kinesins bound vesicles that undergo long‐range transport in neurons. Of these, three had an axonal bias (KIF5B, KIF5C and KIF13B), two were unbiased (KIF1A and KIF1Bβ), and one transported only in dendrites (KIF13A). Overall, the trafficking of vesicle‐bound kinesins to axons or dendrites did not correspond to their motor domain preference, suggesting that on‐vesicle regulation is crucial for kinesin targeting. Surprisingly, several kinesins were associated with populations of somatodendritic vesicles that underwent little long‐range transport. This assay should be broadly applicable for investigating kinesin function in many cell types.

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Section: Resultssupporting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A novel strategy to visualize vesicle‐bound kinesins reveals the diversity of kinesin‐mediated transport

Yang

Bostick

Garbouchian

et al. 2019

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“…To examine the effect of patient variants on the efficiency of KIF1A's microtubule‐based motility, we utilized the neurite tip accumulation assay. In this assay, kinesin motors that are capable of processive microtubule‐based motility accumulate at the tips of neurites in differentiated neuronal cells (Yang, Bentley, Huang, & Banker, 2016). For this assay, we used a truncated and constitutively active version of KIF1A containing amino acids 1–393 tagged with the fluorescent protein monomeric Citrine (mCit) and we expressed wild‐type and mutant versions in differentiated neuronal SH‐SY5Y cells.…”

Section: Resultsmentioning

confidence: 99%

Expansion of the phenotypic spectrum of de novo missense variants in kinesin family member 1A ( KIF1A )

et al. 2020

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Defects in the motor domain of kinesin family member 1A (KIF1A), a neuron‐specific ATP‐dependent anterograde axonal transporter of synaptic cargo, are well‐recognized to cause a spectrum of neurological conditions, commonly known as KIF1A‐associated neurological disorders (KAND). Here, we report one mutation‐negative female with classic Rett syndrome (RTT) harboring a de novo heterozygous novel variant [NP_001230937.1:p.(Asp248Glu)] in the highly conserved motor domain of KIF1A. In addition, three individuals with severe neurodevelopmental disorder along with clinical features overlapping with KAND are also reported carrying de novo heterozygous novel [NP_001230937.1:p.(Cys92Arg) and p.(Pro305Leu)] or previously reported [NP_001230937.1:p.(Thr99Met)] variants in KIF1A. In silico tools predicted these variants to be likely pathogenic, and 3D molecular modeling predicted defective ATP hydrolysis and/or microtubule binding. Using the neurite tip accumulation assay, we demonstrated that all novel KIF1A variants significantly reduced the ability of the motor domain of KIF1A to accumulate along the neurite lengths of differentiated SH‐SY5Y cells. In vitro microtubule gliding assays showed significantly reduced velocities for the variant p.(Asp248Glu) and reduced microtubule binding for the p.(Cys92Arg) and p.(Pro305Leu) variants, suggesting a decreased ability of KIF1A to move along microtubules. Thus, this study further expanded the phenotypic characteristics of KAND individuals with pathogenic variants in the KIF1A motor domain to include clinical features commonly seen in RTT individuals.

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“…Another important kinesin family in neuronal axons consists of the kinesin-3 proteins, including KIF1A and -1B, KIF13A and -13B, and KIF16A and -16B. However, several other kinesins, including KIF2, KIF4, KIF17, and KIF21, are also important in axonal transport (6)(7)(8)(9). Anterograde transport of HSV requires kinesins, but it is currently not clear which kinesins promote this essential process.…”

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confidence: 99%

Kinesin-1 Proteins KIF5A, -5B, and -5C Promote Anterograde Transport of Herpes Simplex Virus Enveloped Virions in Axons

DuRaine

Wisner

Howard

et al. 2018

J Virol

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Herpes simplex virus (HSV) and other alphaherpesviruses must spread from sites of viral latency in sensory ganglia to peripheral tissues, where the viruses can replicate to higher titers before spreading to other hosts. These viruses move in neuronal axons from ganglia to the periphery propelled by kinesin motors moving along microtubules. Two forms of HSV particles undergo this anterograde transport in axons: (i) unenveloped capsids that become enveloped after reaching axon tips and (ii) enveloped virions that are transported within membrane vesicles in axons. Fundamental to understanding this axonal transport is the question of which of many different axonal kinesins convey HSV particles. Knowing which kinesins promote axonal transport would provide clues to the identity of HSV proteins that tether onto kinesins. Prominent among axonal kinesins are the kinesin-1 (KIF5A, -5B, and -5C) and kinesin-3 (e.g., KIF1A and -1B) families. We characterized fluorescent forms of cellular cargo molecules to determine if enveloped HSV particles were present in the vesicles containing these cargos. Kinesin-1 cargo proteins were present in vesicles containing HSV particles, but not kinesin-3 cargos. Fluorescent kinesin-1 protein KIF5C extensively colocalized with HSV particles, while fluorescent kinesin-1 KIF1A did not. Silencing of kinesin-1 proteins KIF5A, -5B, and -5C or light chains KLC1 and KLC2 inhibited the majority of HSV anterograde transport, while silencing of KIF1A had little effect on HSV transport in axons. We concluded that kinesin-1 proteins are important in the anterograde transport of the majority of HSV enveloped virions in neuronal axons and kinesin-3 proteins are less important. Herpes simplex virus (HSV) and other alphaherpesviruses, such as varicella-zoster virus, depend upon the capacity to navigate in neuronal axons. To do this, virus particles tether onto dyneins and kinesins that motor along microtubules from axon tips to neuronal cell bodies (retrograde) or from cell bodies to axon tips (anterograde). Following reactivation from latency, alphaherpesviruses absolutely depend upon anterograde transport of virus particles in axons in order to reinfect peripheral tissues and spread to other hosts. Which of the many axonal kinesins transport HSV in axons is not clear. We characterized fluorescent cellular cargo molecules and kinesins to provide evidence that HSV enveloped particles are ferried by kinesin-1 proteins KIF5A, -5B, and -5C and their light chains, KLC1 and KLC2, in axons. Moreover, we obtained evidence that kinesin-1 proteins are functionally important in anterograde transport of HSV virions by silencing these proteins.

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Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Cited by 11 publications

References 40 publications

A novel strategy to visualize vesicle‐bound kinesins reveals the diversity of kinesin‐mediated transport

A novel strategy to visualize vesicle‐bound kinesins reveals the diversity of kinesin‐mediated transport

Expansion of the phenotypic spectrum of de novo missense variants in kinesin family member 1A ( KIF1A )

Kinesin-1 Proteins KIF5A, -5B, and -5C Promote Anterograde Transport of Herpes Simplex Virus Enveloped Virions in Axons

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