Influence of the 5' noncoding region of hepatitis A virus strain GBM on its growth in different cell lines.
Previous sequence analysis of consecutive passages of the hepatitis A virus (HAV) strain GBM/WT in human embryonic kidney cells (HEK cells), human embryonic lung fibroblasts (HFS cells) and in FRhK-4 cells (foetal rhesus monkey kidney cells) pointed to a host cell dependent cell culture adaptation of GBM/WT in HFS cells involving mutations in the 5' noncoding region (5'NCR). Multiple nucleotide changes occurred in the 5'NCR of the GBM genome after the cell line used for virus passage was changed from HEK cells to HFS cells. In contrast, no mutations in the 5'NCR occurred during the first 20 passages of GBM/WT in FRhK-4 cells. In order to analyse the influence of the 5'NCR on host cell specific adaptation of HAV strain GBM in different cell cultures, GBM/HM175 chimeras were constructed which contained 5'NCRs from different GBM variants by replacing the 5'NCR of the infectious clone pHAV/7. Parallel transfection assays in FRhK-4 and HFS cells, performed with transcripts from the chimeric GBM/HM175 constructs, showed that the 5'NCR of the GBM variant GBM/HFS is essential for virus growth in HFS cells. The GBM/HM175 chimeric RNA, which contained the 5'NCR of GBM/HFS, exclusively, was able to produce infectious virus after transfection of HFS cells. The growth of the different GBM/HM175 chimeras in FRhK-4 cells, in contrast, did not seem to be strongly influenced by a specific sequence of the 5'NCR.
Graff
J
J
Department of Virology and Epidemiology of Virus Diseases, Hygiene Institute, University of Tübingen, Germany. jgraff@uci.edu
Normann
A
A
Flehmig
B
B
D000818:Animals; D002460:Cell Line; D002678:Chimera; D018076:DNA, Complementary / Q000096:biosynthesis; D004622:Embryo, Mammalian; D005347:Fibroblasts; D016679:Genome, Viral; D006507:Hepatovirus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D007668:Kidney; D008168:Lung; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D010957:Plasmids; D012367:RNA, Viral / Q000096:biosynthesis; D014158:Transcription, Genetic; D014162:Transfection
D018076:DNA, Complementary; D012367:RNA, Viral