title: Partial deletion in the spike endodomain of mouse hepatitis virus decreases the cytopathic effect but maintains foreign protein expression in infected cells.
abstract: Mouse hepatitis virus (MHV) produces a series of subgenomic RNAs for viral protein expression. As a prototype coronavirus, MHV has been explored extensively and is often used to express foreign proteins. Previously, a 13-residue deletion in the MHV spike (S) protein endodomain was found to reduce syncytium formation dramatically while inhibiting virus replication slightly. In this study, the effects of the S mutation on MHV infectivity and foreign protein expression were further examined in rat or mouse L2, NIH/3T3 and Neuro-2a cells. The replacement of the MHV 2a/haemagglutinin-esterase gene with a membrane-anchored protein hook (HK) and replacement of gene 4 with EGFP did not change the adaptability and cytopathology of recombinant viruses in these cells. However, the cytopathic effect of the recombinants with the partial S deletion was reduced significantly in these cells. The replication and foreign protein expression of the S-mutated recombinants were found to be more efficient in L2 cells than in Neuro-2a and NIH/3T3 cells. Meanwhile, the distribution patterns of HK and EGFP expressed by the recombinant viruses were similar to those in cells transfected with a eukaryotic expression vector. These results suggest that the partial deletion in the S endodomain may increase the usefulness of MHV as a viral vector by attenuation and maintaining foreign protein expression.
authors.affiliation: Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Yang
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Sun
authors.forename: Yuyan
authors.initials: Y
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Lv
authors.forename: Di
authors.initials: D
authors.lastname: Qu
authors.forename: Rong
authors.initials: R
authors.lastname: Ye
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics* / Q000378:metabolism*; D041681:NIH 3T3 Cells; D010641:Phenotype; D021381:Protein Transport; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins