A Single Point Mutation Creating a Furin Cleavage Site in the Spike Protein Renders Porcine Epidemic Diarrhea Coronavirus Trypsin Independent for Cell Entry and Fusion

Wentao Li; Oliver Wicht; Frank J M van Kuppeveld; Qigai He; Peter J M Rottier; Berend-Jan Bosch

doi:10.1128/JVI.00356-15

A Single Point Mutation Creating a Furin Cleavage Site in the Spike Protein Renders Porcine Epidemic Diarrhea Coronavirus Trypsin Independent for Cell Entry and Fusion

J Virol. 2015 Aug;89(15):8077-81. doi: 10.1128/JVI.00356-15. Epub 2015 May 13.

Authors

Wentao Li¹, Oliver Wicht², Frank J M van Kuppeveld², Qigai He³, Peter J M Rottier², Berend-Jan Bosch⁴

Affiliations

¹ Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, People's Republic of China.
² Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
³ State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, People's Republic of China.
⁴ Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands [email protected].

Abstract

The emerging porcine epidemic diarrhea virus (PEDV) requires trypsin supplementation to activate its S protein for membrane fusion and virus propagation in cell culture. By substitution of a single amino acid in the S protein, we created a recombinant PEDV with an artificial furin protease cleavage site N terminal of the putative fusion peptide (PEDV-SFCS). PEDV-SFCS exhibited trypsin-independent cell-cell fusion and was able to replicate in culture cells independently of trypsin, though to low titer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Amino Acid Substitution
Animals
Coronavirus Infections / enzymology
Coronavirus Infections / veterinary*
Coronavirus Infections / virology
Furin / metabolism*
Point Mutation*
Porcine epidemic diarrhea virus / chemistry
Porcine epidemic diarrhea virus / genetics*
Porcine epidemic diarrhea virus / physiology
Protein Processing, Post-Translational
Spike Glycoprotein, Coronavirus / chemistry
Spike Glycoprotein, Coronavirus / genetics*
Spike Glycoprotein, Coronavirus / metabolism*
Swine
Swine Diseases / enzymology*
Swine Diseases / virology
Trypsin / metabolism*
Virus Internalization*

Substances

Spike Glycoprotein, Coronavirus
Trypsin
Furin