Molecular cloning and functional characterization of porcine nucleotide-binding oligomerization domain-1 (NOD1) recognizing minimum agonists, meso-diaminopimelic acid and meso-lanthionine

Masanori Tohno, Tomoyuki Shimazu, Hisashi Aso, Akiko Uehara, Haruhiko Takada, Akiko Kawasaki, Yukari Fujimoto, Koichi Fukase, Tadao Saito, Haruki Kitazawa

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

In this study, we isolated a complementary DNA encoding nucleotide-binding oligomerization domain-1 (NOD1) from Peyer's patches (Pps) of swine gut-associated lymphoid tissues (GALT). The complete open reading frame of porcine NOD1 contains 2862 bp, encoding a 953-amino acid polypeptide. The porcine NOD1 amino acid sequence is more closely related to the human sequence (83.8% identity) than the mouse counterpart (79.2% identity). To examine the subcellular expression and function of porcine NOD1, we overexpressed it in human embryonic kidney 293 cells. Immunostaining with an anti-porcine NOD1 polyclonal antibody revealed that the protein was expressed in transfectants as an intracellular membrane-bound molecule. In the transfected cells, both γ-d-glutamyl-meso-diaminopimelic acid, and meso-diaminopimelic acid and meso-lanthionine activated nuclear factor-kappa B. Quantitative real-time PCR detected NOD1 mRNA in multiple tissues isolated from adult and newborn swine, including the esophagus, duodenum, jejunum, ileum, ileal Pps, colon, spleen, and mesenteric lymph nodes. In the newborn and adults, NOD1 was highly expressed in the esophagus and GALT, such in the ileal Pps and mesenteric lymph nodes. Furthermore, Toll-like receptor and NOD1 ligands as well as immunobiotic lactic acid bacteria enhanced the expression of NOD1 in GALT of adult and newborn swine. Our results should help clarify how the intestinal immune system is modulated by low-molecular weight peptidoglycan fragments through NOD1.

Original languageEnglish
Pages (from-to)1807-1817
Number of pages11
JournalMolecular Immunology
Volume45
Issue number6
DOIs
Publication statusPublished - 2008 Mar 1
Externally publishedYes

Keywords

  • GALT
  • NOD1
  • Swine
  • cDNA cloning
  • meso-DAP
  • meso-Lanthionine

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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