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An analysis of two open reading frames (ORF3 and ORF4) of rat hepatitis E virus genome using its infectious cDNA clones with mutations in ORF3 or ORF4
- Source :
- Virus Research. 249:16-30
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- Rat hepatitis E virus (ratHEV) genome has four open reading frames (ORFs: ORF1, ORF2, ORF3 and ORF4). The functions of ORF3 and ORF4 are unknown. An infectious cDNA clone (pUC-ratELOMB-131L_wt, wt) and its derivatives including ORF3-defective (ΔORF3) and ORF4-defective (ΔORF4) mutants, were constructed and their full-length RNA transcripts transfected into PLC/PRF/5 cells. ΔORF3 replicated as efficiently as wt in cells. However, ≤1/1000 of the number of progenies were detectable in the culture supernatant of ΔORF3-infected cells compared with wt-infected cells. ORF4 protein was not detectable in ratHEV-infected cells or in the liver tissues of ratHEV-infected rats. No marked differences were noted between wt and ΔORF4 regarding the viral replication and protein expression. ORF3 mutants with proline-to-leucine mutations at amino acids (aa) 93, 96 and/or 98 in ORF3 were constructed and transfected into PLC/PRF/5 cells. Wt and an ORF3 mutant with leucine at aa 98 (ORF3-L98) replicated efficiently (density 1.15–1.16 g/cm3), while ORF3-L93 + L96 exhibited a decreased viral release and banded at 1.26–1.27 g/cm3, similar to ΔORF3. In conclusion, the ORF3 protein, especially its proline residues at aa 93 and 96, is essential for the release of membrane-associated ratHEV particles, and ORF4 is unnecessary for the replication of ratHEV.
- Subjects :
- 0301 basic medicine
Cancer Research
viruses
Mutant
Biology
Virus Replication
medicine.disease_cause
Gene Knockout Techniques
Open Reading Frames
Viral Proteins
03 medical and health sciences
Hepatitis E virus
Virology
Complementary DNA
medicine
Animals
ORFS
RNA
Transfection
Viral Load
Molecular biology
Rats
Open reading frame
030104 developmental biology
Infectious Diseases
Viral replication
Mutant Proteins
Subjects
Details
- ISSN :
- 01681702
- Volume :
- 249
- Database :
- OpenAIRE
- Journal :
- Virus Research
- Accession number :
- edsair.doi.dedup.....a88289168c4a4c42fb3ad4b628f139f1