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Assessing Protein/Protein Interactions Between the Rho GTPase TCL/RhoJ and Coronin Proteins
Author(s) -
Wilson Kaitlyn Guertin,
Tader Brooke R,
Hamann Michael J
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.477.4
Subject(s) - cdc42 , gtpase , prenylation , gtp' , small gtpase , microbiology and biotechnology , fusion protein , rab , vesicular transport protein , biology , vesicle , chemistry , biochemistry , membrane , signal transduction , gene , recombinant dna , enzyme
TCL (RhoJ) is a Rho GTPase that contributes to tumor‐associated angiogenesis and metastatic melanoma. It is part of the Cdc42 subfamily of Rho GTPases, but it has a unique twenty amino acid N terminus that is important for GTP‐loading and localization to the plasma membrane. Deletion of the N terminus promotes GDP‐loading of TCL and its localization to intracellular vesicles. Additional data from our lab has shown that plasma membrane localization of TCL is also dependent on prenylation of the C‐terminal tail, whereas vesicular localization of TCL only required it to be in a GDP‐loaded confirmation. Thus, our hypothesis is that GDP‐loaded TCL is bound to vesicular membranes through a protein/protein interaction. Coronin proteins are logical candidates for this interaction as they bind to other Rho GTPases when they are in a GDP‐loaded conformation. To determine if coronins interact with TCL, the Cdc42‐ and Rac‐interactive binding (CRIB) domains from the seven human coronin isoforms have been PCR amplified and cloned into a pGEX plasmid for GST‐fusion protein purification. The purified CRIB will be used in precipitation assays to test for an interaction with GDP versus GTP loaded TCL. 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 Information This 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). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .