Abstract
Abstract Regulation of MAP Kinase phosphorylation is thought to be important for proper translation of TCR signals into thymocyte cell fates. However, the role of phosphatase activity in thymic development has not been extensively characterized. DUSP-6, also known as Map Kinase Phosphatase-3 (MKP-3), is expressed in the thymus and can bind and dephosphorylate ERK. To explore the role of MKP in thymocytes, we expressed MKP-3 or a catalytically inactive form of MKP-3 (DM-MKP3) in an immature thymocyte cell line, 16610 D9. Lower amounts of phosphorylated ERK (p-ERK) were found in D9 cells that over express MKP-3, while D9 cells that over express DM-MKP3 exhibited higher amounts of p-ERK after TCR stimulation in comparison to vector control cells. Thus, over expression of a catalytically inactive ERK phosphatase results in inhibition of all ERK specific MKP activity, and acts as a dominant negative MKP. Utilizing real time PCR, we also observed elevated Egr-1 mRNA levels after TCR stimulation in D9 cells expressing DM-MKP3 compared to control D9 cells. Egr-1 is an ERK dependent immediate early gene known to support positive selection of thymocytes. Finally, we over expressed DM-MKP3 in vivo, and observed enhanced positive selection. Together these results indicate that MKP control of ERK activation after TCR stimulation is critical in regulation of thymocyte cell fate decisions. This work is funded by grants from the NIH and the ACS.
Published Version
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