Gram-negative bacteria-binding protein, a pattern recognition receptor for lipopolysaccharide and beta-1,3-glucan that mediates the signaling for the induction of innate immune genes in Drosophila melanogaster cells.
Pattern recognition receptors, non-clonal immune proteins recognizing common microbial components, are critical for non-self recognition and the subsequent induction of Rel/NF-kappaB-controlled innate immune genes. However, the molecular identities of such receptors are still obscure. Here, we present data showing that Drosophila possesses at least three cDNAs encoding members of the Gram-negative bacteria-binding protein (DGNBP) family, one of which, DGNBP-1, has been characterized. Western blot, flow cytometric, and confocal laser microscopic analyses demonstrate that DGNBP-1 exists in both a soluble and a glycosylphosphatidylinositol-anchored membrane form in culture medium supernatant and on Drosophila immunocompetent cells, respectively. DGNBP-1 has a high affinity to microbial immune elicitors such as lipopolysaccharide (LPS) and beta-1,3-glucan whereas no binding affinity is detected with peptidoglycan, beta-1,4-glucan, or chitin. Importantly, the overexpression of DGNBP-1 in Drosophila immunocompetent cells enhances LPS- and beta-1,3-glucan-induced innate immune gene (NF-kappaB-dependent antimicrobial peptide gene) expression, which can be specifically blocked by pretreatment with anti-DGNBP-1 antibody. These results suggest that DGNBP-1 functions as a pattern recognition receptor for LPS from Gram-negative bacteria and beta-1, 3-glucan from fungi and plays an important role in non-self recognition and the subsequent immune signal transmission for the induction of antimicrobial peptide genes in the Drosophila innate immune system.
Kim
Y S
YS
Laboratory of Immunology, BK21 Center for Medical Science and Medical Research Center, Yonsei University College of Medicine, 134 Shinchon-dong, CPO Box 8044, Seoul, South Korea.
Ryu
J H
JH
Han
S J
SJ
Choi
K H
KH
Nam
K B
KB
Jang
I H
IH
Lemaitre
B
B
Brey
P T
PT
Lee
W J
WJ
D000209:Acute-Phase Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002352:Carrier Proteins / Q000502:physiology; D002460:Cell Line; D003001:Cloning, Molecular; D004331:Drosophila melanogaster / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D005936:Glucans / Q000378:metabolism*; D019476:Insect Proteins; D008070:Lipopolysaccharides / Q000378:metabolism*; D008668:Metallothionein / Q000235:genetics; D008969:Molecular Sequence Data; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000378:metabolism; D047071:beta-Glucans
D000209:Acute-Phase Proteins; D001798:Blood Proteins; D002352:Carrier Proteins; D005936:Glucans; D019476:Insect Proteins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D047071:beta-Glucans; D008668:Metallothionein; C033363:beta-1,3-glucan; D010738:Type C Phospholipases; D043265:Phosphatidylinositol Diacylglycerol-Lyase