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Ruan, Ying

Cell and Molecular Biology Program

2011

M.S.

Cell migration is a fundamentally important cell behavior which plays a central role in various biological processes. There are various signaling molecules contributable to cell migration, and our lab is focusing on PKA, one of the most prominent kinases involved in signaling network. Previous studies in our lab have shown that PKA is enriched at the leading edge of migrating cells and that it's spatially regulated during chemotactic cell migration. But on how and why PKA getting involved in cell migration is not clearly elucidated yet.
Here, using techniques like two dimensional electrophoresis and mass spectrometry on analyzing the protrusive part of the cell during cell migration, we identified a new substrate for PKA: 14-3-3[epsilon]. As the most conserved member in the 14-3-3 protein family, 14-3-3[epsilon] has shown its interactions with many other signaling molecules involved in cell migration. Additionally, sequence screen for 14-3-3[epsilon] showed that it contains a very strong PM site, RRAS. Kinase assay for GST-tagged 14-3-3[epsilon] showed that the bacterial 14-3-3[epsilon] can be phosphorylated by PKA, while transfected cells treated with PKA activators and inhibitors showed that the rat isoform of 14-3-3[epsilon] can be phosphorylated by endogenous PKA.