title: Novel myeloid differentiation factor 88, EsMyD88, exhibits EsTube-binding activity in Chinese mitten crab Eriocheir sinensis.
abstract: Myeloid differentiation factor 88 (MyD88) is a universal and essential adapter protein that participates in the activation of the Toll-like receptor/interleukin-1 receptor-mediated signaling pathway. In the present study, a new MyD88 gene (named EsMyD88) was identified in the Chinese mitten crab Eriocheir sinensis. The cDNA of EsMyD88 was 2210 bp long with a 1416 bp open reading frame that encoded a protein with 472 amino acids. Predicted EsMyD88 protein had a death domain at the N-terminal and a TIR domain at the C-terminal. BLASTP and phylogenetic analysis results showed that EsMyD88 was clustered in one group together with other crustaceans MyD88 (SpMyD88, FcMyD88, LvMyD88, and LvMyD88-1). EsMyD88 was detected in all the examined tissues of healthy crabs, and was mainly expressed in the hemocytes and nerves. When normal crabs were challenged with lipopolysaccharide, peptidoglycan, Staphylococcus aureus, Vibrio parahaemolyticus, or Aeromonas hydrophila, the expression levels of EsMyD88 significantly increased either in the hepatopancreas or hemocytes. Results of the pull-down assay showed that EsMyD88 could bind to downstream cytosolic adaptor EsTube. Overexpression of EsMyD88 protein in Drosophila Schneider 2 cells led to the activation of antimicrobial peptide genes. RNA interference assay showed that EsMyD88 is involved in regulating the transcription of ALF1 and ALF2, Cru1 and Cru2, and Lys in crab challenged with V. parahaemolyticus. All the results mentioned earlier indicated that EsMyD88 gene has a key function in antibacterial innate immune defense.
authors.affiliation: Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.affiliation: MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory of Biocontrol, School of Marine Sciences, Sun Yat-sen University, Guangzhou, China. Electronic address: chenyihong365@gmail.com.
authors.forename: Yi-Hong
authors.initials: YH
authors.lastname: Chen
authors.affiliation: Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China. Electronic address: njnuwang@263.net.
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China. Electronic address: renqian0402@126.com.
authors.forename: Qian
authors.initials: Q
authors.lastname: Ren
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000276:immunology*; D016980:Aeromonas hydrophila / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000276:immunology; D060829:Arthropod Proteins / Q000235:genetics / Q000276:immunology*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000276:immunology*; D003386:Brachyura / Q000187:drug effects / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000276:immunology; D006434:Hemocytes / Q000187:drug effects / Q000276:immunology / Q000382:microbiology; D043143:Hepatopancreas / Q000187:drug effects / Q000276:immunology / Q000382:microbiology; D054884:Host-Pathogen Interactions; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000276:immunology*; D016366:Open Reading Frames; D010802:Phylogeny; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013211:Staphylococcus aureus / Q000276:immunology; D014736:Vibrio parahaemolyticus / Q000276:immunology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D051792:Basic Helix-Loop-Helix Proteins; D008070:Lipopolysaccharides; D053594:Myeloid Differentiation Factor 88
keywords: Antibacterial peptides; Binding activity; Eriocheir sinensis; EsMyD88; Innate immunity