Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8751816 | Virus Research | 2018 | 50 Pages |
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 (ÎORF3) and ORF4-defective (ÎORF4) mutants, were constructed and their full-length RNA transcripts transfected into PLC/PRF/5 cells. ÎORF3 replicated as efficiently as wt in cells. However, â¤1/1000 of the number of progenies were detectable in the culture supernatant of ÎORF3-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 ÎORF4 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 1.15-1.16â¯g/cm3), while ORF3-L93â¯+â¯L96 exhibited a decreased viral release and banded at 1.26-1.27â¯g/cm3, similar to ÎORF3. 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.
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Authors
Tanggis Tanggis, Tominari Kobayashi, Masaharu Takahashi, Suljid Jirintai, Tsutomu Nishizawa, Shigeo Nagashima, Takashi Nishiyama, Satoshi Kunita, Emiko Hayama, Takeshi Tanaka, Mulyanto Mulyanto, Hiroaki Okamoto,