Loss-of-function phenotype of a PKCθT219A knockin mouse strain

Autor: Nikolaus Thuille, Kerstin Siegmund, Victoria Klepsch, Jacqueline Schörgenhuber, Sarah Danklmaier, Michael Leitges, Gottfried Baier
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Cell Communication and Signaling, Vol 17, Iss 1, Pp 1-11 (2019)
Druh dokumentu: article
ISSN: 1478-811X
DOI: 10.1186/s12964-019-0466-8
Popis: Abstract Background Protein kinase C θ has been established as an important signaling intermediate in T-effector-cell activation and survival pathways by controlling activity of the key transcription factors NF-κB and NFAT. Previous studies identified an activation-induced auto-phosphorylation site at Thr-219, located between the tandem C1 domains of the regulatory fragment in PKCθ, as a structural requirement for its correct membrane translocation and the subsequent transactivation of downstream signals leading to IL-2 production in a human T cell line. Methods The present work aimed to define the role of this phosphorylation switch on PKCθ in a physiological context through a homozygous T219A knockin mouse strain. T cell activation was analyzed by H3-thymidine uptake (proliferative response), qRT-PCR and luminex measurements (cytokine production). NFAT and NF-κB transactivation responses were estimated by Gel mobility shift and Alpha Screen assays. Frequencies of T cell subsets were analyzed by flow cytometry. Results Despite a normal T cell development, in vitro activated effector T cells clearly revealed a requirement of Thr-219 phosphorylation site on PKCθ for a transactivation of NF-κB and NFAT transcription factors and, subsequently, robust IL-2 and IFN-γ expression. Conclusion This phenotype is reminiscent of the PKCθ knockout T cells, physiologically validating that this (p) Thr-219 auto-phosphorylation site indeed critically regulates PKCθ function in primary mouse T cells.
Databáze: Directory of Open Access Journals
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