Screening and identification of SipC-interacting proteins in Salmonella enteritidis using Gal4 yeast two-hybrid system in duck

PeerJ

article

Yu Zhang¹  Tiantian Gu¹  Yang Chen¹  Guoqiang Zhu¹  Wanwipa Vongsangnak³  Qi Xu¹  Guohong Chen¹ 

[1] Joint International Research Laboratory of Agriculture & Agri-Product Safety of Ministry of Education, Yangzhou University;Jiangsu Key Laboratory of Zoonosis, Yangzhou University;Department of Zoology, Faculty of Science, Kasetsart University

关键词: Duck;    PERP;    TAB2;    Salmonella enteritidis;    SipC protein;   

DOI  :  10.7717/peerj.7663

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

The zoonotic pathogen Salmonella not only reduces the production performance in ducks, but also poses a serious threat to human health through eggs and pollutes water bodies through feces. SipC, an effector protein of type III secretion systems (T3SS) in Salmonella, mediates translocation of effectors into the eukaryotic host. However, the precise role of SipC effectors remains unknown in ducks. In this study, the SipC from duck granulosa cells (dGCs) was selected as bait, and the SipC-interacting proteins in Salmonella enteritidis (SE) were screened using Gal4 yeast two-hybrid system in duck. Twelve SipC-interacting proteins were identified. Among those, the p53-effector related to PMP-22 (PERP) and TGF-β activated kinase 1-binding protein 2 (TAB2) were selected to further confirm the function by GST pull-down in vitro. Over-expression of PERP resulted in not only increasing SE adhesion and invasion but also triggering the production of IL-1β and IFN-α in SE infected dGCs, while knock-down PERP showed the opposite tendency (P < 0.01). In addition, TAB2 significantly induced the production of IL-6, IL-1β, IFN-α, and INF-γ in SE infected dGCs (P < 0.05), but did not cause obvious changes in SE adhesion and invasion. When the sipC in SE was deleted, the activities of duck PERP and TAB2 were abolished because they could not bind to SipC. Taken together, although the protein of PERP and TAB2 can interact with SipC, their mechanisms were different in duck challenged by SE. Therefore, PERP was involved in SE invasion and inflammatory response of dGC ovaries, and TAB2 only contributed to dGCs inflammatory response, which provided critical insights about the mechanism in host- bacterium protein interactions during Salmonella invasion in duck.

【 授权许可】

CC BY