A systematic analysis of the effects of splicing on the diversity of post-translational modifications in protein isoforms.

Post-translational modifications (PTMs) and splicing are known to be important regulatory processes for controlling protein function and activity. However, there have been limitations in analyzing the interplay of alternative splicing and PTMs, which stems from the deep differences in genomic and proteomic databases. In this work, we bridged the protein- and genome-centric world views to map PTMs to genomic locations for subsequent projection of PTMs onto alternative isoforms. We then performed a systematic analysis of the diversification of PTMs by alternative splicing, including exploration of the modification-specific rates of inclusion across isoforms and how often the regulatory sequences directly flanking a PTM are impacted by splicing, which might indicate altered regulatory or binding interactions in the alternatively spliced isoform. We found that 6-51% of PTMs are excluded from at least one isoform, depending on the modification type. Further, approximately 2% of prospective PTM sites exhibited altered regulatory sequences surrounding the modification site, suggesting that regulatory or binding interactions might be diversified in these proteoforms. Lastly, we applied this PTM-to-isoform mapping approach to explore the impacts of disease-related splicing in prostate cancer, identifying possible new hypotheses explaining the variable consequences of ESRP1 expression in different cancers. As a part of this work, we have provided an easily implementable tool for annotating splice events identified from RNA-sequencing with PTMs and their functional consequences, called PTM-POSE.