Analyzing the role of U2AF1 isoforms on the activation of the inflammatory pathway
Alternative splicing is a crucial process that diversifies gene expression and increases organismal diversity. Defects in alternative splicing can significantly impact various biological pathways, particularly the inflammatory pathway, leading to the development of conditions such as cancer, myelodysplastic syndromes, and genomic changes. One of the most important proteins involved in alternative splicing is U2AF1, which recognizes and binds to the 3' splice site of an intron, allowing the cleaving system to know where to begin the removal of an intron.
The expression of different isoforms of U2AF1 (a and b) can lead to slight alterations in the binding site of U2AF1, as well as varying levels of binding at different 3' splice sites. These changes can subsequently impact a regulatory pathway in protein production, thereby altering cellular responses. Investigations on the cellular outcomes of U2AF1 isoform-exclusive cell lines under diverse extracellular stresses have revealed that the inflammatory pathway, particularly the NF-kB pathway, experiences drastic changes in gene expression.
The NF-kB pathway plays a crucial role in both the initiation and resolution phases of inflammation. This pathway will amplify the inflammatory response and support recovery in multiple ways. The NF-kB pathway remains dormant unless inflammation occurs, but when inflammation occurs, the NF-kB pathway becomes active. The protein p65, which is a part of the NF-kB complex, is the primary protein that will be utilized to activate various immune response genes. The activated form of p65, known as p-P65, will be measured to understand the level of immune activation in the different isoform-expressed cell lines.