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The C3 domain of Pasteurella multocida toxin is the minimal domain responsible for activation of G q ‐dependent calcium and mitogenic signaling
Author(s) -
Aminova Leila R.,
Luo Shuhong,
Bannai Yuka,
Ho Mengfei,
Wilson Brenda A.
Publication year - 2008
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.083445408
Subject(s) - nfat , luciferase , signal transduction , calcineurin , biology , calmodulin , pasteurella multocida , microbiology and biotechnology , amino acid , biochemistry , transfection , transcription factor , enzyme , genetics , gene , bacteria , medicine , surgery , transplantation
Abstract The large 1285‐amino‐acid protein toxin from Pasteurella multocida (PMT) is a multifunctional single‐chain polypeptide that binds to and enters eukaryotic cells and acts intracellularly to promote G q and G 12/13 protein‐dependent calcium and mitogenic signal transduction. Previous studies indicated that the intracellular activity domain responsible for PMT action was located within the C‐terminal 600–700 amino acids. In this study, we have exogenously expressed a series of N‐ and C‐terminal PMT fragments directly in mammalian cells and have used the dual luciferase reporter system to assay for toxin‐mediated activation of calcium‐calcineurin‐NFAT signaling (NFAT‐luciferase) and mitogenic serum response signaling (SRE‐luciferase). Using this approach, we have defined the last 180 amino acids, which encompass the C3 domain in the crystal structure, as the minimum domain sufficient to activate both NFAT and SRE signaling pathways.