Original ResearchBasic and Translational—Alimentary TractTLE1 Modifies the Effects of NOD2 in the Pathogenesis of Crohn's Disease
Section snippets
Y2H Screen
The Clontech MATCHMAKER Gal4 Two-hybrid System 3 (Takara Bio, St Germain en Laye, France) was used for the identification of NOD2 interacting partners based on standard methodology.20 A human intestinal cell line (SW480) complementary DNA (cDNA) library was constructed in pGADT7-AD. Full-length human NOD2 cDNA was generated from RNA isolated from SW480 cells and cloned, in frame, into the yeast expression vector pGBKT7-BD. NOD2 in SW480 contains the P268S variant (rs2066842). This was altered
Y2H Screen
The Y2H screen was performed using full-length NOD2 as the bait and a cDNA library constructed from RNA extracted from SW480 cells. TLE1, HIV-1 Tat interactive protein (HTATIP), protein phosphatase 2, regulatory subunit B′, epsilon isoform (PPP2R5E), fission 1 (mitochondrial outer membrane) homolog (FIS1), UDP-N-acetyl-alpha-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2 (GALNT2), and Vimentin were identified as potential interacting proteins with NOD2 (Table 3).
Discussion
By using a Y2H screen we have identified a number of proteins interacting with NOD2 as potential candidate genes for IBD susceptibility. None of the genes had been associated previously with NOD2, and thus each discovery implicates novel pathways in which NOD2 is involved. With the exception of the GALNT2 SNPs, none of these genes were identified in any of the published genome-wide association studies or meta-analyses of genome-wide association studies to date.26 HTATIP, PPP2R5E, FIS1, and
Acknowledgments
The authors thank Gemma Williams for analysis of preliminary genotyping data; the Wellcome Trust Clinical Research Facility for Illumina Goldengate genotyping, TaqMan genotyping, and DNA extraction; and the MRC Human Genetics Unit for DNA extraction and sequencing.
References (40)
Clinical epidemiology of inflammatory bowel disease: incidence, prevalence, and environmental influences
Gastroenterology
(2004)- et al.
TNF-[alpha] and IFN-[gamma] regulate the expression of the NOD2 (CARD15) gene in human intestinal epithelial cells
Gastroenterology
(2003) - et al.
CARD15/NOD2 functions as an antibacterial factor in human intestinal epithelial cells
Gastroenterology
(2003) - et al.
NOD2/CARD15 mediates induction of the antimicrobial peptide human beta-defensin-2
J Biol Chem
(2006) - et al.
A role for Erbin in the regulation of Nod2-dependent NF-kappaB signaling
J Biol Chem
(2005) - et al.
Reciprocal cross-talk between Nod2 and TAK1 signaling pathways
J Biol Chem
(2004) - et al.
GRIM-19 interacts with nucleotide oligomerization domain 2 and serves as downstream effector of anti-bacterial function in intestinal epithelial cells
J Biol Chem
(2005) - et al.
Centaurin beta-1 down-regulates nucleotide-binding oligomerization domains 1- and 2-dependent NF-kappaB activation
J Biol Chem
(2006) - et al.
Mammalian Ras interacts directly with the serine/threonine kinase Raf
Cell
(1993) - et al.
Tip60 Is a Co-repressor for STAT3
J Biol Chem
(2003)
Cellular integrity plus: organelle-related and protein-targeting functions of intermediate filaments
Trends Cell Biol
A role for Erbin in the regulation of Nod2-dependent NF-kappaB signaling
J Biol Chem
GRIM-19 interacts with nucleotide oligomerization domain 2 and serves as downstream effector of anti-bacterial function in intestinal epithelial cells
J Biol Chem
Genetics of inflammatory bowel disease: progress and prospects
Hum Mol Genet
The prevalence of inflammatory bowel disease in relatives of patients with Crohn's disease
Eur J Gastroenterol Hepatol
The genetics of Crohn's disease
Ann Rev Genomics Human Genet
Genetics of inflammatory bowel disease: implications for disease pathogenesis and natural history
Exp Rev Gastroenterol Hepatol
Genome-wide meta-analysis increases to 71 the number of confirmed Crohn's disease susceptibility loci
Nat Genet
Meta-analysis identifies 29 additional ulcerative colitis risk loci, increasing the number of confirmed associations to 47
Nat Genet
Genome-wide association study of ulcerative colitis identifies three new susceptibility loci, including the HNF4A region
Nat Genet
Cited by (32)
Gut permeability among Astronauts during Space missions
2024, Life Sciences in Space ResearchNegative regulation of human mononuclear phagocyte function
2013, Mucosal ImmunologyRoles of transducin-like enhancer of split (TLE) family proteins in tumorigenesis and immune regulation
2022, Frontiers in Cell and Developmental BiologyWeighted gene co-expression network analysis identifies dysregulated B-cell receptor signaling pathway and novel genes in pulmonary arterial hypertension
2022, Frontiers in Cardiovascular MedicineA Pharmacoinformatics Analysis of Artemisinin Targets and de novo Design of Hits for Treating Ulcerative Colitis
2022, Frontiers in Pharmacology
Conflicts of interest The authors disclose no conflicts.
Funding Supported by a Wellcome Trust (Molecular and Cell Biology Panel) program grant (072789/Z/03/Z) and grants from the Chief Scientist Office (Scotland) (CZB/4/146 and CZB/4/585); and by an MRC patient cohort research initiative grant (G0800675).