ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/060/SRR13775760/SRR13775760_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/062/SRR13775762/SRR13775762_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/056/SRR13775756/SRR13775756_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/059/SRR13775759/SRR13775759_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/061/SRR13775761/SRR13775761_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/062/SRR13775762/SRR13775762_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/058/SRR13775758/SRR13775758_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/059/SRR13775759/SRR13775759_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/063/SRR13775763/SRR13775763_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/064/SRR13775764/SRR13775764_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/057/SRR13775757/SRR13775757_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/056/SRR13775756/SRR13775756_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/057/SRR13775757/SRR13775757_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/058/SRR13775758/SRR13775758_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/063/SRR13775763/SRR13775763_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/064/SRR13775764/SRR13775764_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/061/SRR13775761/SRR13775761_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR137/060/SRR13775760/SRR13775760_1.fastq.gzprimaryOK200GenomicsDepartment of Life Sciences, University of Triestehttps://www.ebi.ac.uk/ena/browser/view/PRJNA704548Homo sapiensxref:PubMed:35835219An emerging area of TDP-43 is represented by the study of post-translational modifications and the way they are connected to disease-associated mutations. Recently, we described a novel mutation in TDP-43 in an early-onset ALS case that was affecting a potential phosphorylation site in position 375 (S375G). A preliminary characterization showed that both the S375G variant and its phosphomimic mutant, S375E, displayed altered nuclear-cytoplasmic distribution and cellular toxicity. To better investigate these effects, we established cell lines expressing inducible TDP-43 WT, S375G, and S375E variants. No significant changes were reported in splicing of known regulated targets, TDP-43 autoregulation, or aggregation. However, cell-cycle analysis of the stable clones showed that the number of cells in the G2 phase decreased in the two phospho-mutants (S375G and S375E), as compared to WT. Furthermore, experiments showed that Apoptosis-inducing factor 1 (AIF1) appeared to be released from the mitochondria. The involvement of apoptosis and alterations in cell-cycle induced by these variants were further validated using RNAseq analysis. Taken together, these data strongly support the growing evidence that alterations of TDP-43 post-translational modifications can play a potentially important role in disease pathogenesis. Overall design: Generation of Tet-inducible Hek293 Flp-In T-REx monoclones expressing Wild-Type, S375G, and S375E variants of human TDP-43.ENAfalseAlterations in cell cycle progression caused by S375G and S375E phosphomutants of TDP-43Alterations in cell cycle progression caused by S375G and S375E phosphomutants of TDP-432025-09-242022-10-07PRJNA704548GSE167385960635835219