HuD regulates SOD1 expression during oxidative stress in differentiated neuroblastoma cells and sporadic ALS motor cortex

The database includes the raw of the article “HuD regulates SOD1 expression during oxidative stress in differentiated neuroblastoma cells and sporadic ALS motor cortex”.

In detail, the database contains data obtained by the following evaluations: i) RNA electrophoretic mobility shift assay (REMSA) to detect the formation of a complex between HuD recombinant protein and SOD1 ARE sequences. ii) multiplex qRT-PCR to evaluate if oxidative stress levels, i.e H2O2 treatment, induces a shift in the alternative polyadenylation (APA) site usage in SOD1 3'UTR.

Aim of the work was to investigate the potential involvement of HuD (ELAVL4) one of the neuronal members of the ELAVL family, in SOD1 regulation during oxidative stress and in sporadic ALS patients (sALS).

Using differentiated SH-SY5Y cells along with brain tissues from sALS patients, we evaluated HuD-dependent regulation of SOD1 mRNA. By means in vitro binding and mRNA decay assays we observed that HuD specifically binds to SOD1 ARE motifs promoting mRNA stabilization. In SH-SY5Y cells, we faound that the overexpression of full-length HuD increased SOD1 mRNA and protein levels; moreover HuD regulation of SOD1 mRNA we found that this regulation is related to oxidative stress, as shown after H2O2 exposure. H2O2 exposure also induced a shift in the APA site usage in SOD1 3'UTR, increasing the levels of a long variant bearing HuD binding sites. We validated this data using a specific siRNA. In the motor cortex from sALS patients, we found increases in SOD1 and HuD mRNAs and proteins, accompanied by greater HuD binding to this mRNA as confirmed by RNA-immunoprecipitation (RIP) assays.

To conclude, these results point a role of HuD in the post-transcriptional regulation of SOD1 expression in neurons after oxidative stress and in sALS brain tissues, thus suggesting an involvement in ALS pathogenesis.