Doctoral thesis (Dissertations and theses)
Interaction between M protein and HSC70 directs clathrin-mediated endocytosis of transmissible gastroenteritis virus
Dong, Hui
2019
 

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Keywords :
TGEV; M protein; HSC70
Abstract :
[en] TGEV infection has been reported in many countries around the world, causing highly contagious enteric disease in piglets. TGEV causes severe diarrhea, vomiting and dehydration in the piglets. The mortality rates of TGEV infection are high in pigs of all ages, reaching up to 100% in piglets less than 2 weeks old. Moreover the recovered piglets can also spread the virus to uninfected pig for many weeks. My thesis project aims to characterize a coronavirus membrane (M) protein. In particular the TGEV M protein acts as a major player in virus assembly and possibly interacts with host factors to modulate cellular functions. In the first part of my thesis, we prepared the monoclonal antibodies to TGEV N protein and identify the epitopes on TGEV N protein. The mAb (5E8) can be applied for different approaches to diagnosis of TGEV infection. In the second part of my thesis, we prepared the monoclonal antibodies to TGEV M protein and identify the epitopes on TGEV M protein. The mAb (1C3) can be used for the detection of the TGEV M protein in different assays. An indirect immunofluorescence assay (IFA) method for detection of TGEV M protein was optimized using mAb (1C3). In the third part of my thesis, cellular proteins interacting with TGEV M protein were screened by endogenous Co-IP coupled with MALDI-TOF/TOF MS. The cellular protein HSC70 was identified to interact with TGEV M directly. The endodomain of TGEV M and substrate binding domain (SBD) of HSC70 are essential for the interaction. HSC70 downregulation, as well as HSC70 ATPase activity inhibition, suppresses the TGEV replication. TGEV enters into the PK-15 cells by clathrin-mediated endocytosis, and interaction between M protein and HSC70 directs clathrin-mediated endocytosis of TGEV. In summary, my thesis contributed to a better understanding of TGEV infection.
Research center :
Laboratory of Cellular and Molecular Epigenetics (University of Liege)
Harbin Veterinary Research Institute (Chinese Academy of Agricultural Sciences)
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Dong, Hui ;  Université de Liège - ULiège > Doct. sc. agro. & ingé. biol. (Paysage)
Language :
English
Title :
Interaction between M protein and HSC70 directs clathrin-mediated endocytosis of transmissible gastroenteritis virus
Defense date :
13 June 2019
Institution :
ULiège - Université de Liège
Degree :
doctorate in agronomy and bioengineering
Funding text :
2015BAD12B02, The National Key Research and Development Program of China; 2016YFD0500103, The National Key Research and Development Program of China; 2017YFD0500104, The National Key Research and Development Program of China; 31572541, National Natural Science Foundation of China; 31502092, National Natural Science Foundation of China; SKLVBP2015005, the National Key Laboratory of the Veterinary Biotechnology Support Program of China
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since 06 June 2019

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