A β-integrin from sea cucumber Apostichopus japonicus exhibits LPS binding activity and negatively regulates coelomocyte apoptosis. (May 2016)

• Record Type: Journal Article
• Authors: Wang, Zhenhui; Shao, Yina; Li, Chenghua; Lv, Zhimeng; Wang, Haihong; Zhang, Weiwei; Zhao, Xuelin
• Is Part Of: Fish & shellfish immunology. Volume 52(2016:Sep.)
• Page Start: 103
• Page End: 110
• Publication Date: 2016-05
• Subjects: Apostichopus japonicus -- β-Integrin -- LPS binding activity -- Cell apoptosis -- siRNA; cDNA Complementary DNA -- RACE Rapid-amplification of cDNA ends -- RGD The tripeptide Arg-Gly-Asp -- LPS Lipopolysaccharide -- PGN Peptidoglycan -- MAN Mannotriose-di-(N-acetyl-d-glucosamine) -- SUS Skin ulceration syndrome -- qPCR Quantitative PCR -- PAMPs Pathogen-associated molecular patterns -- ERK Extracellular regulated protein kinases -- GAS Group A Streptococcus -- SDS-PAGE SDS-polyacrylamide gel electrophoresis; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2016.03.031
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Integrins are a family of membrane glycoproteins, which are the major receptors for extracellular matrix and cell–cell adhesion molecules. In this study, a 1038 bp sequence representing the full-length cDNA of a novel β-integrin subunit (designated as AjITGB ) was cloned from Apostichopus japonicus by using combined transcriptome sequencing and RACE approaches. The deduced amino acid sequence of AjITGB shared a conserved tripeptide Arg-Gly-Asp (RGD) binding domain with an S-diglyceridecysteine or N-Palm cysteine residue (C 31 ), a transmembrane domain, and a β-integrin cytoplasmic domain. Spatial distribution analysis showed that AjITGB was constitutively expressed in all tested tissues with dominant expression in the muscles and weak expression in the respiratory tree. The pathogen Vibrio splendidus challenge and LPS stimulation could both significantly down-regulate the mRNA expression of AjITGB . Functional investigation revealed that recombinant AjITGB displayed higher LPS binding activity but lower binding activity to PGN and MAN. More importantly, knockdown of AjITGB by specific siRNA resulted in the significant promotion of coelomocyte apoptosis in vitro . Results indicated that AjITGB may serve as an apoptosis inhibitor with LPS binding activity during host–pathogen interaction in sea cucumber. Highlights: Full-length cDNA of integrin-β subunit were identified in Apostichopus japonicus. AjIGTB were ubiquitously expressed in all examined tissues. The V. splendidus challenge and LPS exposure could both depress AjIGTB mRNA expression. Recombinant AjITGB showed high LPS binding activities and lower to PGN and MAN. Silencing the AjITGB promoted coelomocytes apoptosis in vitro.