Caenorhabditis elegans dnj-14, the orthologue of the DNAJC5 gene mutated in adult onset neuronal ceroid lipofuscinosis, provides a new platform for neuroprotective drug screening and identifies a SIR-2.1-independent action of resveratrol

Kashyap, Sudhanva S.; Johnson, James R.; McCue, Hannah V.; Chen, Xi; Edmonds, Matthew J.; Ayala, Mimieveshiofuo; Graham, Margaret E.; Jenn, Robert C.; Barclay, Jeff W.; Burgoyne, Robert D.; Morgan, Alan

doi:10.1093/hmg/ddu316

Cited by 44 publications

(65 citation statements)

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“…For example, a spontaneous genetic mutation that robustly delays synaptic degeneration (known as Wld S ) can ameliorate neurodegeneration across a range of conditions, including some forms of motor neuron disease (Ferri et al 2003), Parkinson's disease (Sajadi et al 2004) and following global cerebral ischemia (Gillingwater et al 2004) or traumatic brain injury . Furthermore, at the molecular level, a number of individual synaptic proteins that can modulate synaptic vulnerability and degeneration in response to a variety of pathological stimuli have recently been identified, including CSP alpha/DNAJC5 (Fern andez-Chac on Garc ıa-Junco-Clemente et al 2010;Noskov a et al 2011;Wishart et al 2012;Kashyap et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Molecular neuropathology of the synapse in sheep with CLN5 Batten disease

et al. 2015

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AimsSynapses represent a major pathological target across a broad range of neurodegenerative conditions. Recent studies addressing molecular mechanisms regulating synaptic vulnerability and degeneration have relied heavily on invertebrate and mouse models. Whether similar molecular neuropathological changes underpin synaptic breakdown in large animal models and in human patients with neurodegenerative disease remains unclear. We therefore investigated whether molecular regulators of synaptic pathophysiology, previously identified in Drosophila and mouse models, are similarly present and modified in the brain of sheep with CLN5 Batten disease.MethodsGross neuropathological analysis of CLN5 Batten disease sheep and controls was used alongside postmortem MRI imaging to identify affected brain regions. Synaptosome preparations were then generated and quantitative fluorescent Western blotting used to determine and compare levels of synaptic proteins.ResultsThe cortex was particularly affected by regional neurodegeneration and synaptic loss in CLN5 sheep, whilst the cerebellum was relatively spared. Quantitative assessment of the protein content of synaptosome preparations revealed significant changes in levels of seven out of eight synaptic neurodegeneration proteins investigated in the motor cortex, but not cerebellum, of CLN5 sheep (α‐synuclein, CSP‐α, neurofascin, ROCK2, calretinin, SIRT2, and UBR4).ConclusionsSynaptic pathology is a robust correlate of region‐specific neurodegeneration in the brain of CLN5 sheep, driven by molecular pathways similar to those reported in Drosophila and rodent models. Thus, large animal models, such as sheep, represent ideal translational systems to develop and test therapeutics aimed at delaying or halting synaptic pathology for a range of human neurodegenerative conditions.

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Section: Introductionmentioning

confidence: 99%

Molecular neuropathology of the synapse in sheep with CLN5 Batten disease

et al. 2015

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“…For example, mutation of a Drosophila PAT, HIP14, causes a strong impairment of neurotransmission that is entirely due to non-palmitoylation of cysteine string protein (CSP), as expression of a chimaeric CSP artificially targeted to synapses fully rescues this defect [ 59 ]. However, the extremely severe phenotype of CSP mutants seen in Drosophila [ 60 ] is not observed in C. elegans null mutants [ 61 ], so even if mutation of a worm PAT(s) completely prevented CSP palmitoylation, this would not result in strong effects on the phenotypes analysed in the present study.…”

Section: Discussionmentioning

confidence: 84%

A systematic analysis of protein palmitoylation in Caenorhabditis elegans

Edmonds

Morgan

2014

BMC Genomics

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BackgroundPalmitoylation is a reversible post-translational protein modification which involves the addition of palmitate to cysteine residues. Palmitoylation is catalysed by the DHHC family of palmitoyl-acyl transferases (PATs) and reversibility is conferred by palmitoyl-protein thioesterases (PPTs). Mutations in genes encoding both classes of enzymes are associated with human diseases, notably neurological disorders, underlining their importance. Despite the pivotal role of yeast studies in discovering PATs, palmitoylation has not been studied in the key animal model Caenorhabditis elegans.ResultsAnalysis of the C. elegans genome identified fifteen PATs, using the DHHC cysteine-rich domain, and two PPTs, by homology. The twelve uncategorised PATs were officially named using a dhhc-x system. Genomic data on these palmitoylation enzymes and those in yeast, Drosophila and humans was collated and analysed to predict properties and relationships in C. elegans. All available C. elegans strains containing a mutation in a palmitoylation enzyme were analysed and a complete library of RNA interference (RNAi) feeding plasmids against PAT or PPT genes was generated. To test for possible redundancy, double RNAi was performed against selected closely related PATs and both PPTs. Animals were screened for phenotypes including size, longevity and sensory and motor neuronal functions. Although some significant differences were observed with individual mutants or RNAi treatment, in general there was little impact on these phenotypes, suggesting that genetic buffering exists within the palmitoylation network in worms.ConclusionsThis study reports the first characterisation of palmitoylation in C. elegans using both in silico and in vivo approaches, and opens up this key model organism for further detailed study of palmitoylation in future.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-841) contains supplementary material, which is available to authorized users.

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“…Moreover, the absence of CSPα leads to neurodegeneration in drosophila, mice and c. elegans 20,41,77 . In vitro, DnaJB6 inhibits Aβ amyloid aggregation 47 .…”

Section: Discussionmentioning

confidence: 99%

“…We found that the molecular co-chaperone, CSPα (cysteine string protein, DnaJC5) promotes EV export of distinct misfolded disease-causing proteins and that this export pathway is 'druggable' as demonstrated by the ability of the polyphenol, resveratrol, to block CSPα-mediated export of misfolded proteins 15 . Resveratrol was initially linked to CSPα function in a chemical screen as a compound that alters life span of CSPα c. elegans mutants 41 .…”

Section: Introductionmentioning

confidence: 99%

“…Loss-of-function CSPα Drosophila mutants demonstrate uncoordinated movements, temperature-sensitive paralysis and early lethality 77 . And, in C elegans, CSPα null mutants display neurodegeneration and reduced lifespan 42 . Not surprisingly, CSPα dysfunction has been implicated in several neurodegenerative disorders in addition to ANCL, including Alzheimer's, Parkinson's and Huntington's disease 7,16,34,50,69,76 .…”

Section: Introductionmentioning

confidence: 99%

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Cargo-Loading of Misfolded Proteins into Extracellular Vesicles: The CSPα-EV Export Pathway

Pink

Donnelier

Lewis

et al. 2018

Preprint

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Extracellular vesicles (EVs) are a collection of secreted vesicles of diverse size and cargo that are implicated in the physiological removal of nonfunctional proteins as well as the cell-tocell transmission of disease-causing-proteins in several neurodegenerative diseases. We have shown that the molecular chaperone, cysteine string protein (CSPα; DnaJC5), is responsible for the export of disease-causing-misfolded proteins from neurons via EVs. We show here that CSPα-EVs efficiently deliver GFP-tagged 72Q huntingtin exon1 to naive neurons. When we analyzed the heterogeneous EV pool, we found that the misfolded GFP-tagged 72Q huntingtin exon1 cargo was primarily found in EVs between 180-240nm. We further determined that cargo-loading of GFP-tagged 72Q huntingtin exon1 into EVs was impaired by resveratrol. Importantly, in addition to CSPα, we identified two other J protein co-chaperones, DnaJB2 and DnaJB6, that facilitate EV export of GFP-tagged 72Q huntingtin exon1 . While human mutations in CSPα cause the neurodegenerative disorder, adult neuronal ceroid lipofuscinosis, mutations in DnaJB6 cause limb-girdle muscular dystrophy and mutations in DnaJB2 are linked to the neurodegenerative disorders, Charcot Marie Tooth disease, distal hereditary motor neuropathy, spinal muscular atrophy and juvenile Parkinsonism. Our data provides new insights into the parallels between proteostasis and EV export, as three J proteins linked to disease in humans are the same as those that mediate EV genesis and export of misfolded proteins.CSPα (DnaJC5) 1-198 DnaJB2 1-324-long DnaJB6 1-365-long J J CCC J CAAX box UIM UIM NLS

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Caenorhabditis elegans dnj-14, the orthologue of the DNAJC5 gene mutated in adult onset neuronal ceroid lipofuscinosis, provides a new platform for neuroprotective drug screening and identifies a SIR-2.1-independent action of resveratrol

Cited by 44 publications

References 54 publications

Molecular neuropathology of the synapse in sheep with CLN5 Batten disease

Molecular neuropathology of the synapse in sheep with CLN5 Batten disease

A systematic analysis of protein palmitoylation in Caenorhabditis elegans

Cargo-Loading of Misfolded Proteins into Extracellular Vesicles: The CSPα-EV Export Pathway

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