Investigating the Phosphorylation of Mnk2 Isoforms by p38 MAPK as a Potential Cancer Therapy Target
Abstract (300 words maximum)
A critical cell signaling pathway, known as the p38 MAPK pathway, responds to various extracellular stimuli such as inflammation and stress. Downstream effects upon activation of p38 MAPK lead to the phosphorylation of other proteins including the mitogen-activated protein kinase signal-integrating kinase Mnk2, which is involved in cellular process regulation of the proliferation of cells and protein synthesis. Mnk2 proteins have been included in cancer research studies due to their potential role in tumor growth and have been shown in the regulation of gene expression for the cell cycle and apoptosis. Studies have demonstrated that inhibition of Mnk2 leads to a suppression of tumor growth and therefore a cancer therapy target. Of the two known isoforms, Mnk2a acts as a tumor suppressor and is downregulated in some cancerous tumors, and Mnk2b has been found to be pro-oncogenic. The isoforms are splice variants and most studies have only focussed on isoform a. Very little is known about how isoform b functions. Previous research has been conducted demonstrating the phosphorylation of Mnk2a by p38 kinase, however the phosphorylation of Mnk2b remains unknown. To better understand the function of the two isoforms of Mnk2, in this project, we determine the phosphorylation of the isoforms by activated p38. This is an ongoing project conducted as a course-based undergraduate research experience (CURE) for the undergraduate course Chem 3512L. His-tagged MNK isoform proteins were purified using a nickel column. Two isoform specific antibodies were tested by western blotting and showed that the antibodies are able to distinguish between the isoforms. With this data, the CURE project will be concluded by conducting kinase assays using purified his-tagged Mnk2 isoforms and isoform specific antibodies.The phosphorylation of the two isoforms will be detected using western blotting.Results in this study will be the first on phosphorylation and potential activation of MNK isoform b.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Rajnish Singh
Investigating the Phosphorylation of Mnk2 Isoforms by p38 MAPK as a Potential Cancer Therapy Target
A critical cell signaling pathway, known as the p38 MAPK pathway, responds to various extracellular stimuli such as inflammation and stress. Downstream effects upon activation of p38 MAPK lead to the phosphorylation of other proteins including the mitogen-activated protein kinase signal-integrating kinase Mnk2, which is involved in cellular process regulation of the proliferation of cells and protein synthesis. Mnk2 proteins have been included in cancer research studies due to their potential role in tumor growth and have been shown in the regulation of gene expression for the cell cycle and apoptosis. Studies have demonstrated that inhibition of Mnk2 leads to a suppression of tumor growth and therefore a cancer therapy target. Of the two known isoforms, Mnk2a acts as a tumor suppressor and is downregulated in some cancerous tumors, and Mnk2b has been found to be pro-oncogenic. The isoforms are splice variants and most studies have only focussed on isoform a. Very little is known about how isoform b functions. Previous research has been conducted demonstrating the phosphorylation of Mnk2a by p38 kinase, however the phosphorylation of Mnk2b remains unknown. To better understand the function of the two isoforms of Mnk2, in this project, we determine the phosphorylation of the isoforms by activated p38. This is an ongoing project conducted as a course-based undergraduate research experience (CURE) for the undergraduate course Chem 3512L. His-tagged MNK isoform proteins were purified using a nickel column. Two isoform specific antibodies were tested by western blotting and showed that the antibodies are able to distinguish between the isoforms. With this data, the CURE project will be concluded by conducting kinase assays using purified his-tagged Mnk2 isoforms and isoform specific antibodies.The phosphorylation of the two isoforms will be detected using western blotting.Results in this study will be the first on phosphorylation and potential activation of MNK isoform b.