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Effects of the NOD2 Mutations E600A, T605P, and T605N on NF‐κB Activation
Author(s) -
Domina Aaron,
Stiles Ethan,
Ewer Roger,
Bocian Charlie,
Lee Soojin,
Saab Mohamad
Publication year - 2015
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.29.1_supplement.763.3
Subject(s) - nod2 , muramyl dipeptide , nod , transfection , mutant , mutation , hek 293 cells , pyrin domain , biology , microbiology and biotechnology , genetics , gene , innate immune system , receptor , inflammasome , immune system
Mutations in the NOD2 gene predispose individuals to Crohn's Disease, Blau Syndrome, familial uveitis, and other diseases depending on the location of the mutation. NOD2 is a pattern recognition receptor for muramyl dipeptide (MDP), a component of the bacterial peptidoglycan layer, as well as single stranded viral RNA. In most cases of Blau Syndrome, the mutations are found within the NOD/NACHT domain, which promotes NOD2 oligomerization, resulting in constitutive NF‐kB activation; however Blau syndrome and familial uveitis‐associated mutations outside the NOD/NACHT domain have not been well‐characterized. In this current study, three disease‐associated mutations that are found outside of the NOD/NACHT domain were created (E600A, T605P, and T605N) and compared to wild‐type (WT) NOD2, as well as to a NOD/NACHT domain mutation commonly found in Blau Syndrome (R334W). The objective of this study was to determine whether the non‐NOD/NACHT mutants (E600A, T605P, and T605N) activate NF‐kB in a similar manner as the R334W mutation. To this end, each mutant, WT NOD2, and a vector control were separately transfected into human embryonic kidney cells‐293 (HEK‐293). Transfected cells were incubated with or without MDP, followed by lysate preparation and Western blot analysis for NF‐kB activation. The results of this work provide insight into the effects of these non‐NOD/NACHT mutations on the NF‐kB pathway and suggest a means for therapeutic intervention. This work was supported by grants from Husson University and the Husson University School of Pharmacy.