Formation of retromer transport carriers is disrupted by the Parkinson disease‐linked Vps35<scp>D620N</scp>variant

Cui, Yi; Yang, Zhe; Flores‐Rodriguez, Neftali; Follett, Jordan; Ariotti, Nicholas; Wall, Adam A.; Parton, Robert G.; Teasdale, Rohan D.

doi:10.1111/tra.12779

Cited by 27 publications

(40 citation statements)

References 75 publications

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“…Our live cell imaging revealed that retromer endosomes labeled by Vps35D620N were less dynamic than those labeled by wildtype Vps35 and detected less frequently small, motile, Tetherin-containing vesicles in cells expressing mutant Vps35 with D620N than in cells expressing wildtype Vps35. These observations are consistent with recent findings showing that Vps35D620N impairs the production of transport carriers [21].…”

Section: Discussionsupporting

confidence: 94%

“…The D620N mutation in Vps35 does not affect the formation of the Vps26/Vps35/Vps29 heterodimer nor alter cargo recognition [17]. Ectopically expressed as well as endogenous Vps35 with the D620N mutation induces the enlargement of retromer-positive endosomes and impairs the formation of retromer transport carriers and retrieval of dopamine D1 receptors, dopamine transporters, and many other proteins, including those involved in the degradation of α-synuclein [17][18][19][20][21]. The presence of D620N in Vps35 also impairs autophagy and causes accumulation of PD-variant α-synuclein A53T [22].…”

Section: Introductionmentioning

confidence: 99%

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Parkinson’s Disease Causative Mutation in Vps35 Disturbs Tetherin Trafficking to Cell Surfaces and Facilitates Virus Spread

Ding

Chhetri

et al. 2021

Cells

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Parkinson’s disease (PD) is the most common neurodegenerative movement disorder, characterized by progressive loss of dopaminergic neurons in the substantia nigra, intraneuronal deposition of misfolded proteins known as Lewy bodies, and chronic neuroinflammation. PD can arise from monogenic mutations, but in most cases, the etiology is unclear. Viral infection is gaining increasing attentions as a trigger of PD. In this study, we investigated whether the PD-causative 620 aspartate (D) to asparagine (N) mutation in the vacuolar protein sorting 35 ortholog (Vps35) precipitated herpes simplex virus (HSV) infection. We observed that ectopic expression of Vps35 significantly reduced the proliferation and release of HSV-1 virions; the D620N mutation rendered Vps35 a partial loss of such inhibitory effects. Tetherin is a host cell protein capable of restricting the spread of encapsulated viruses including HSV-1 and SARS-Cov-2, both of which are implicated in the development of parkinsonism. Compared with cells overexpressing wildtype Vps35, cells expressing mutant Vps35 with D620N had less Tetherin on cell surfaces. Real-time and static cell imaging revealed that Tetherin recycled through Vps35-positive endosomes. Expression of Vps35 with D620N reduced endosomal dynamics and frequency of motile Tetherin-containing vesicles, a sign of defective production of recycling carriers. Our study suggests that the D620N mutation in Vps35 hinders Tetherin trafficking to cell surfaces and facilitates virus spread.

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Section: Discussionsupporting

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Parkinson’s Disease Causative Mutation in Vps35 Disturbs Tetherin Trafficking to Cell Surfaces and Facilitates Virus Spread

Ding

Chhetri

et al. 2021

Cells

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“…1A.i-ii; 1-way ANOVA p = 0.44). In cell lines, the D620N mutation impairs VPS35 association with the Wiskott-Aldrich syndrome protein and SCAR homolog (WASH) complex member family with sequence similarity 21 (FAM21) by coIP (44,45,47). We found the level of FAM21 pulled by VPS35 was similarly reduced in coIP of whole-brain lysates from mutant mice (Fig.…”

Section: Altered Protein Binding Relationships and Lrrk2 Kinase Activity In Vki Brainmentioning

confidence: 76%

“…Previous reports of VPS35 neurobiology at glutamatergic synapses, and the effects of the D620N mutant, have relied on exogenous protein expression or knock-out, and produced somewhat con icting results. Studies on retrograde tra cking of the canonical retromer cargo cation-independent mannose-6phosphate receptor (CI-MPR), for example, concluded that the mutation results in a loss-of-function (15,44,46,47) or has no effect (45,70). This discordance may result from the variety of cell types and modes of expression used to glean insights.…”

Section: Discussionmentioning

confidence: 99%

Endosomal Traffic And Glutamate Synapse Activity Are Increased In VPS35 D620N Mutant Knock-In Mouse Neurons, And Resistant To LRRK2 Kinase Inhibition

Kadgien

Kamesh

Khinda

et al. 2021

Preprint

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Vacuolar protein sorting 35 (VPS35) regulates neurotransmitter receptor recycling from endosomes. A missense mutation (D620N) in VPS35 leads to autosomal-dominant, late-onset Parkinson’s disease. Here, we study the basic neurobiology of VPS35 and Parkinson’s disease mutation effects in the D620N knock-in mouse and the effect of leucine-rich repeat kinase 2 (LRRK2) inhibition on synaptic phenotypes. The study was conducted using a VPS35 D620N knock-in mouse that expresses VPS35 at endogenous levels. Protein levels, phosphorylation states, and binding ratios in brain lysates from knock-in mice and wild-type littermates were assayed by co-immunoprecipitation and western blot. Dendritic protein co-localization, AMPA receptor surface expression, synapse density, and glutamatergic synapse activity in primary cortical cultures from knock-in and wild-type littermates were assayed using immunocytochemistry and whole-cell patch clamp electrophysiology. In brain tissue, we confirm VPS35 forms complexes with LRRK2 and AMPA-type glutamate receptor GluA1 subunits, in addition to NMDA-type glutamate receptor GluN1 subunits and D2-type dopamine receptors. Receptor and LRRK2 binding was unaltered in D620N knock-in mice, but we confirm the mutation results in reduced binding of VPS35 with WASH complex member FAM21, and increases phosphorylation of the LRRK2 kinase substrate Rab10, which is reversed by LRRK2 kinase inhibition in vivo. In cultured cortical neurons from knock-in mice, pRab10 is also increased, and reversed by LRRK2 inhibition. The mutation also results in increased endosomal recycling protein cluster density (VPS35-FAM21 co-clusters and Rab11 clusters), glutamate transmission, and GluA1 surface expression. LRRK2 kinase inhibition, which reversed Rab10 hyper-phosphorylation, did not rescue elevated glutamate release or surface GluA1 expression in knock-in neurons, but did alter AMPAR traffic in wild-type cells. The results improve our understanding of the cell biology of VPS35, and the consequences of the D620N mutation in developing neuronal networks. Together the data support a chronic synaptopathy model for latent neurodegeneration, providing phenotypes and candidate pathophysiological stresses that may drive eventual transition to late-stage parkinsonism in VPS35 PD. The study demonstrates the VPS35 mutation has effects that are independent of ongoing LRRK2 kinase activity, and that LRRK2 kinase inhibition alters basal physiology of glutamate synapses in vitro.

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“…Previous reports of VPS35 neurobiology at glutamatergic synapses, and the effects of the D620N mutant, have relied on exogenous protein expression or knock-out, and produced somewhat conflicting results. Studies on retrograde trafficking of the canonical retromer cargo cation-independent mannose-6-phosphate receptor (CI-MPR), for example, concluded that the mutation results in a loss-of-function (15,44,46,47) or has no effect (45,70). This discordance may result from the variety of cell types and modes of expression used to glean insights.…”

Section: Discussionmentioning

confidence: 99%

Endosomal traffic and glutamate synapse activity are increased in VPS35 D620N mutant knock-in mouse neurons, and resistant to LRRK2 kinase inhibition

Kadgien

Kamesh

Khinda

et al. 2021

Preprint

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Background Vacuolar protein sorting 35 (VPS35) regulates neurotransmitter receptor recycling from endosomes. A missense mutation (D620N) in VPS35 leads to autosomal-dominant, late-onset Parkinson’s disease. Methods Here, we study the basic neurobiology of VPS35 and Parkinson’s disease mutation effects in the D620N knock-in mouse and the effect of leucine-rich repeat kinase 2 (LRRK2) inhibition on synaptic phenotypes. The study was conducted using a VPS35 D620N knock-in mouse that expresses VPS35 at endogenous levels. Protein levels, phosphorylation states, and binding ratios in brain lysates from knock-in mice and wild-type littermates were assayed by co-immunoprecipitation and western blot. Dendritic protein co-localization, AMPA receptor surface expression, synapse density, and glutamatergic synapse activity in primary cortical cultures from knock-in and wild-type littermates were assayed using immunocytochemistry and whole-cell patch clamp electrophysiology. Results In brain tissue, we confirm VPS35 forms complexes with LRRK2 and AMPA-type glutamate receptor GluA1 subunits, in addition to NMDA-type glutamate receptor GluN1 subunits and D2-type dopamine receptors. Receptor and LRRK2 binding was unaltered in D620N knock-in mice, but we confirm the mutation results in reduced binding of VPS35 with WASH complex member FAM21, and increases phosphorylation of the LRRK2 kinase substrate Rab10, which is reversed by LRRK2 kinase inhibition in vivo. In cultured cortical neurons from knock-in mice, pRab10 is also increased, and reversed by LRRK2 inhibition. The mutation also results in increased endosomal recycling protein cluster density (VPS35-FAM21 co-clusters and Rab11 clusters), glutamate transmission, and GluA1 surface expression. LRRK2 kinase inhibition, which reversed Rab10 hyper-phosphorylation, did not rescue elevated glutamate release or surface GluA1 expression in knock-in neurons, but did alter AMPAR traffic in wild-type cells. Conclusions The results improve our understanding of the cell biology of VPS35, and the consequences of the D620N mutation in developing neuronal networks. Together the data support a chronic synaptopathy model for latent neurodegeneration, providing phenotypes and candidate pathophysiological stresses that may drive eventual transition to late-stage parkinsonism in VPS35 PD. The study demonstrates the VPS35 mutation has effects that are independent of ongoing LRRK2 kinase activity, and that LRRK2 kinase inhibition alters basal physiology of glutamate synapses in vitro.

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Formation of retromer transport carriers is disrupted by the Parkinson disease‐linked Vps35D620Nvariant

Cited by 27 publications

References 75 publications

Parkinson’s Disease Causative Mutation in Vps35 Disturbs Tetherin Trafficking to Cell Surfaces and Facilitates Virus Spread

Parkinson’s Disease Causative Mutation in Vps35 Disturbs Tetherin Trafficking to Cell Surfaces and Facilitates Virus Spread

Endosomal Traffic And Glutamate Synapse Activity Are Increased In VPS35 D620N Mutant Knock-In Mouse Neurons, And Resistant To LRRK2 Kinase Inhibition

Endosomal traffic and glutamate synapse activity are increased in VPS35 D620N mutant knock-in mouse neurons, and resistant to LRRK2 kinase inhibition

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