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The C‐terminal Tail of TCL Localizes the GTPase to the Plasma Membrane of HeLa Cells
Author(s) -
Tader Brooke,
Florke Rebecca R,
Hamann Michael J
Publication year - 2017
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.31.1_supplement.619.19
Subject(s) - gtpase , subcellular localization , cdc42 , n terminus , microbiology and biotechnology , gtp' , small gtpase , vesicle , c terminus , peptide sequence , amino acid , biology , biochemistry , chemistry , membrane , gene , cytoplasm , signal transduction , enzyme
TCL/RhoJ belongs to the Rho family of GTPases, and it has been shown to contribute to angiogenesis and tumor growth. To better understand the inherent biochemistry of TCL, our lab has been investigating primary sequence difference between TCL and its closest homologs Cdc42 and TC10. Thus far, amino acids 17–20 in the N terminus have been found to be important for GTP‐loading and localization of TCL to the plasma membrane, and mutation of this sequence leads to TCL localization to vesicles. To better understand how the TCL N‐terminus, C‐terminus, and GTP‐binding regions contribute to membrane and vesicle localization independently of each other, expression constructs were generated that fuse the first 24 and last 21 amino acids of TCL to the fluorescent protein Venus. Interestingly, these constructs indicate that the C‐terminus alone may be mostly responsible for TCL plasma membrane localization. TCL constructs containing the GTPase core without the C‐terminal tail suggest the GTPase region uniquely controls localization to vesicles. Additional mutations in the C‐terminal tail will be generated to test how the C‐terminal CAAX sequence and the polybasic sequence contribute to TCL localization in order to understand how TCL localization regulates its function.