Identifying Direct Interactions Between the Rho GTPase TCL and Coronin CRIB Motifs

Abstract

TCL is a Rho GTPase that contributes to tumor‐associated angiogenesis and metastatic melanoma; however, less is known about its basic cell biology and biochemistry. Currently, our lab has found that GTP‐loaded TCL tends to localize to the plasma membrane, while GDP‐loaded TCL localizes to intracellular vesicles. Our data also suggests that vesicle localization of GDP‐loaded TCL occurs through a protein/protein interaction. Coronins are a logical partner for this interaction as they bind to other Rho GTPases when they are in a GDP‐loaded form. To test for a direct interaction between Coronins and TCL, the CRIB domains from the seven human Coronin isoforms have been cloned into plasmids for use in interaction assays using both traditional GST‐fusion protein precipitation and protein complementation using an in vitro split luciferase assay system. Given the importance of TCL in angiogenesis and melanoma, delineation of its interactions with partner proteins is essential for understanding its role in pathology‐related processes.Support or Funding InformationThis work was supported by Bemidji State University Biology Department and the College of Business, Mathematics, and Science. Support was also provided through the Neilson Foundation, Bemidji, MN and Regenerative Medicine Minnesota (RMM‐2017‐EP‐04).