A Preliminary Study On Identification And Regulatory Mechanism Of MAP3K8 Intron Retention

Alternative splicing of pre-m RNA is an important post-transcriptional regulatory mechanism that is ubiquitous in eukaryotes,including humans.Alternative splicing is playing an important role in the process of improving the protein diversity,regulating cell growth,differentiation and development,and maintaining the normal life activities of the body.Among the types of alternative splicing that have been reported,there are few studies on intron retention.Our early analysis found that the intron retention phenomenon of MAP3K8 gene,in view of the important function of this gene in immunity,inflammation and tumor,this paper intends to verify the intron retention of MAP3K8 gene and its regulatory mechanism through experiments.This study identifies the retention of intron 2 of the gene MAP3K8 and verifies that the MAP3K8 transcript retained intron is located in the nucleus.In order to further explore the regulatory mechanism of MAP3K8 intron retention,we construct a series of recombinant plasmids and find that there may be a cis-regulatory element in MAP3K8 retained intron to inhibit the intron splicing.Through si RNA knockdown experiments,it is found that PTBP1 may inhibit the splicing of intron of gene MAP3K8.In order to verify whether the retention of intron transcript in the nucleus is to regulate the production and expression of MAP3K8 in response to extracellular stimuli or signal changes.LPS is used as its stimulating factor,and intron retention is detected by different LPS concentration gradient treatments.The result shows that the intron retention of MAP3K8 is changed relative to the control after LPS treatment,but there is no significance.In addition,we also find that the full-length and truncated MAP3K8 proteins are located in cell differently,and it is speculated that their functions have changed.Our research has laid a certain foundation for exploring the regulatory mechanism of intron retention,and provids new ideas for the in-depth understanding of the function of the gene MAP3K8.