Investigating the effects of mutations of amino acids on the protein expression of CDK2 cancer gene

Each year, twenty million people are diagnosed with cancer. Its prevalence is increasing at a faster pace than any other disease. Cyclin-Dependent Kinase 2 (CDK2) plays a key role in regulating the cell cycle. When CDK2 is overexpressed, it disrupts the cell cycle by accelerating transitions and bypassing checkpoints, leading to uncontrolled cell growth, which is characterized as a hallmark of cancer. There are two approaches to address CDK2 overexpression: one option entails creating mutations in the CDK2 gene that immobilizes its functionality, effectively stopping the cell from dividing and keeping it in a resting phase (G0 phase). The alternative option is to use CDK2 inhibitors that neutralize its activity. We hypothesized that creating mutations in CDK2 gene’s crucial amino acids would lead to significant changes in protein production. We selected seven strategic mutations in two essential regions: the glycine-rich loop and the activation loop. Mutation E12R, located in the glycine-rich loop, inhibited CDK2 gene expression by 80% compared to wild type, while mutation T160E-E162T, located in the activation loop, showed 95% inhibition. Mutation T160L, located in the activation loop, exhibited protein expression levels similar to wild type, whereas mutation T160E, located in the activation loop, resulted in a four-fold increase in CDK2 protein content. The results show that mutations E12R and T160E-E162T were most effective at suppressing CDK2 protein concentration; however, mutation T160E had an opposite effect, leading to increased CDK2 protein concentration. These findings highlight the importance of crucial amino acids in CDK2 expression and their impact on cell growth.