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Structural recognition and stabilization of tyrosine hydroxylase by the J-domain protein DNAJC12

ABSTRACT: Pathogenic variants of the J-domain protein DNAJC12, a co-chaperone of Hsp70, cause parkinsonism, which seems associated with a defective interaction of DNAJC12 with tyrosine hydroxylase (TH), the rate limiting enzyme in dopamine synthesis. Here, we report the characterization of TH:DNAJC12 complex formation and show that DNAJC12 binding stabilizes TH and delays its time-dependent aggregation in an Hsp70-independent manner, while maintaining TH activity and regulatory inhibition by dopamine. Interestingly, although neither TH nor DNAJC12 alone significantly [MDT1] [AM2] activate Hsc70, the complex stimulates [AM3] its ATPase activity. Cryoelectron microscopy reveals two DNAJC12 monomers bound per TH tetramer, each embracing one of the two regulatory domain dimers, leaving all active sites available for substrate and dopamine interaction. Biochemical data confirm the key role of the DNAJC12 last eight residues in TH binding, explaining the molecular disease mechanism of C-terminal truncated DNAJC12 variants.

INSTRUMENT(S): Orbitrap Exploris 240

ORGANISM(S): Homo Sapiens (human)

DISEASE(S): Neurodegenerative Disease

SUBMITTER: Jimena Muntaner

LAB HEAD: José María

PROVIDER: PXD045185 | Pride | 2023-12-01

REPOSITORIES: Pride

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Project description:Action of alcohol on synaptic mRNA in the amygdala of mice Chronic alcohol consumption induces changes in gene expression, causing persistent long-term neuro-adaptations and the remodeling of synaptic structures. These alcohol-induced synaptic changes may rely specifically on the local translation of mRNAs in the synaptic compartments of the cell. We profiled the transcriptome from synaptoneurosomes (SN) and paired total homogenates (TH) of amygdala to analyze the synaptic adaptations induced by chronic voluntary alcohol consumption in mice. In the SN both the number of alcohol-responsive mRNAs and the magnitude of fold-change were greater than in the TH. Accordingly, the SN detected many genes with coordinated patterns of expression producing a highly connected mRNA network in gene co-expression analysis. The greater sensitivity of the SN preparation allowed for improved cell-type specificity analysis, revealing an up-regulation of alcohol-responsive astrocytic and microglial modules that correlated with alcohol consumption. Alcohol was found to induce changes in the SN functionally important biological pathways, including long-term potentiation, long-term potentiation depression, glutamate pathway, neuro-immune, RNA-processing and translational machineries and provided overlap with changes seen in human alcoholic brain. Transposable elements were responsive to alcohol and found in the down-regulated neuronal mRNA module, which may underlie some of the coordinated gene expression changes associated with alcohol. We provide evidence that enrichment of synaptic components reveals a more intricate network of coordinated gene expression. Increased resolution captures the molecular effects of synaptic manipulations and provides an improved technique for identifying therapeutic targets for alcohol abuse. Synaptoneurosomes (SN) and Paired total homogenates (TH) were prepared from the same homogenate. 42 microarrays were used in total for the alcohol-control analysis (8 alcohol treated mice and 13 controls), 21 SN and 21 paired TH. 2 TH (control) samples were found outliers and were removed from the analysis. For the SN vs. TH analysis, the 2 TH samples found outliers were removed with the 2 paired SN samples.

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Structural recognition and stabilization of tyrosine hydroxylase by the J-domain protein DNAJC12

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