Expression profiling by high throughput sequencing
Summary
Global protein synthesis is upregulated in cancers, and thus cancer cells are prone to errors in translation, which may lead to protein misfolding and proteotoxic stress. The pathways that maintain protein homeostasis support the transformation and growth of cancer cells. Here, we show that the cytosolic NAD+ synthesis pathway (controlled by NMNAT-2) plays an essential role in ovarian cancer by maintaining protein homeostasis by regulating the activity of PARP-16, that MARylates the ribosomal proteins. Using polysome-RNA Seq analysis, we found that inhibiting ribosomal protein MARylation by knocking down NMNAT-2 or PARP-16 leads to increased association of specific mRNAs with polysomes and enhanced translation of a set of mRNAs that contain specific secondary structures in their 3’UTRs, which leads to protein aggregation. Collectively, our study demonstrates that NMNAT-2 regulates PARP-16 activity and ribosomal protein MARylation, which control protein homeostasis by fine-tuning the levels of protein synthesis in ovarian cancers.
Overall design
To evaluate the changes in mRNA loading on polysomes when NMNAT-2 or PARP-16 are knockdown, we performed polysome-RNA-Seq analysis on these cells. We isolated total RNA (Input RNA) and polysome-associated RNA from OVCAR3 cells subjected to NMNAT2 or PARP16 knockdown and performed RNA-Seq analysis. Please note that each processed data was generated from both replicates and is linked to the corresponding replicate1 sample records.
Less...
More...