title: Spiroplasma eriocheirisAdhesin-Like Protein (ALP) Interacts with Epidermal Growth Factor (EGF) Domain Proteins to Facilitate Infection.
abstract: Spiroplasma eriocheirisis a novel pathogen found in recent years, causing the tremor disease (TD) of Chinese mitten crabEriocheir sinensis. LikeSpiroplasma mirum, S. eriocheirisinfects the newborn mouse (adult mice are not infected) and can cause cataract. Adhesion-related protein is an important protein involved in the interaction between pathogen and host. In this study, the Adhesin-like Protein (ALP) ofS. eriocheiriswas detected on its outer membrane by using immune electron microscopy, and was found to be involved in the bacterium's infection of mouse embryo fibroblasts (3T6-Swiss albino). Yeast two-hybrid analysis demonstrated that ALP interacts with a diverse group of mouse proteins. The interactions between recombinant partial fibulin7 (FBLN7; including two epidermal growth factor [EGF] domains) and ALP were confirmed by Far-western blotting and colocalization. We synthetized the domains of FBLN7 [EGF domain: amino acids 136-172 and complement control protein (CCP) domain: 81-134 amino acids], and demonstrated that only EGF domain of FBLN7 can interact with ALP. Because the EGF domain has high degree of similarity to EGF, it can activate the downstream EGFR signaling pathway, in key site amino acids. The EGFR pathway in 3T6 cells was restrained after rALP stimulation resulting from competitive binding of ALP to EGF. The unborn mouse, newborn mouse, and the adult mouse with cataract have a small amount of expressed FBLN7; however, none was detected in the brain and very little expression was seen in the eye of normal adult mice. In short, ALP as aS. eriocheirissurface protein, is critical for infection and further supports the role of ALP inS. eriocheirisinfection by competitive effection of the EGF/EGFR axis of the target cells.
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Libo
authors.initials: L
authors.lastname: Hou
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Gao
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Xuechuan
authors.initials: X
authors.lastname: Xu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Mingxiao
authors.initials: M
authors.lastname: Ning
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Jingxiu
authors.initials: J
authors.lastname: Bi
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal UniversityNanjing, China; Co-Innovation Center for Marine Bio-Industry Technology of Jiangsu ProvinceLianYungang, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University Nanjing, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal UniversityNanjing, China; Co-Innovation Center for Marine Bio-Industry Technology of Jiangsu ProvinceLianYungang, China.
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Meng
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004705:Endocytosis; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D005347:Fibroblasts / Q000382:microbiology; D054884:Host-Pathogen Interactions; D051379:Mice; D016253:Microscopy, Immunoelectron; D011485:Protein Binding; D025941:Protein Interaction Mapping; D015398:Signal Transduction; D013149:Spiroplasma / Q000737:chemistry / Q000502:physiology*; D020798:Two-Hybrid System Techniques
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002135:Calcium-Binding Proteins; C060820:fibulin; D004815:Epidermal Growth Factor; C541485:EGFR protein, mouse; D066246:ErbB Receptors
keywords: 3T6-Swiss albino; Spiroplasma eriocheiris; adhesin-like protein; epidermal growth factor; fibulin7