Identification of HSPA5 targets involved in non-alcoholic fatty liver disease

Background: It has been reported that HSPA5 is an endoplasmic reticulum chaperone, which regulates cell metabolism, especially lipid metabolism. There are many reports about the interaction between HSPA5 and protein regulating cell function, but there is no report about HSPA5 binding RNA and its biological function. However, it may have RNA binding ability by using interaction capture. Method: In the present study, the ability of HSPA5 to modulate the AS of cellular genes was assessed using a high-throughput RT-PCR approach to examine AS in lipid metabolism-associated genes. RNA immunoprecipitation coupled to RNA sequencing (RIP-Seq) assays were also performed to identify cellular mRNA sassessment. Results: Upon HSPA5 expression, we detected modifications to the AS profiles of numerou genes involved in pathology of nonalcoholic fatty liver disease. Moreover, we show that HSPA5 modulates the expression levels of various splicing factors such as NEAT1, LRP1, and EGFR. Finally, RNA immunoprecipitation coupled to RIP-Seq assays demonstrate that HSPA5 immunoprecipitates specific cellular mRNAs. Conclusion: The HSPA5 protein can modulate the AS profiles of numerous cellular genes and it binds lncRNA and mRNA, and the related genes are links to nonalcoholic fatty liver disease, which is also the innovation of this project. RIP-seq analysis of HSPA5 IP versus input in Hela cells.