FUNCTIONAL ANALYSIS OF REGULATORY ELEMENTS IN SIGNAL TRANSDUCTION PATHWAYS

Abstract

Regulatory DNA elements including promoters, enhancers and silencers are significant and play a role in regulation of gene expression in cellular systems and are closely coordinated with Signal transduction pathways, enabling cells to dynamically respond to both internal and external stimuli. Although there has been a great breakthrough in molecular biology, the functional nature of many of the regulatory elements in signaling networks has not been fully determined. The purpose of this study was the identification and functional characterization of important regulatory factors in the important signaling pathways as well as the effect of these factors on the activity of pathways and the expression of genes. They used a mix of both computational and experimental methods, i.e. sequence analysis and motif prediction to identify regulatory elements, which were then functionally validated by reporter gene assays and site-directed mutagenesis. Changes in gene expression of the treatment were measured by qPCR and pathway interactions were analyzed using established signaling databases and protein interaction networks with relevant statistical analysis conducted to guarantee significance. The findings indicated the presence of a number of putative regulatory factors of the signaling pathway genes with functional assays showing that the one or the other enhancer and promoter elements affect the expression of the genes greatly. The importance of these regulatory elements in pathway regulation was further established by mutational analysis, which showed that pathway activity can be adjusted by changes in these regulatory elements. In sum, the evidence presented in the study indicates that regulatory factors are critical regulating factors of signal transduction pathways that would enrich our understanding of how genes should be regulated and provide valuable information that could be used in future studies in molecular genetics and development of therapeutic targets.