The cytopathic 18f strain of Hepatitis A virus induces RNA degradation in FrhK4 cells.
The mechanism responsible for the induction of apoptosis by the rapidly replicating HM175/18f strain of Hepatitis A virus (HAV) was investigated. Full length HAV RNA and viral capsid protein VP1 were detected in 18f infected cells at earlier times post-infection than in HM175/clone 1 infected cells. Analysis of total cellular RNA from HM175/18f infected FrhK4 cells by denaturing agarose gel electrophoresis and Northern blot hybridization revealed extensive degradation of both the 28S and 18S ribosomal RNA (rRNA) molecules. Similar degradation was observed when these cells were infected with Human coxsackievirus B1, a fast replicating enterovirus. In contrast, the parental strain of 18f, HM175/clone 1 did not induce RNA degradation. Inhibition of RNA degradation correlated with inhibition of virus replication. The pattern of rRNA degradation resembled degradation of rRNAs by RNase L, an enzyme activated in interferon-treated cells following infection with certain viruses. Ribosomal RNA degradation was accompanied by the reduction in the levels of several cellular RNAs including those for beta-actin and glyceraldehyde-3-phosphate dehydrogenase, while the levels of c-myc and c-jun were higher. Interferon mRNAs could not be detected in either infected or mock-infected control cells, and STAT1, a key regulator of interferon action was not phosphorylated following virus infection. These results reveal a heretofore-undescribed pathway that involves the regulation of RNA degradation and apoptosis following HAV/18f replication in FrhK4 cells.
Kulka
M
M
Food and Drug Administration, Center for Food Safety and Applied Nutrition, Division of Molecular Biology, Laurel, MD 20708, USA.
Chen
A
A
Ngo
D
D
Bhattacharya
S S
SS
Cebula
T A
TA
Goswami
B B
BB
D000818:Animals; D015152:Blotting, Northern; D020126:Brefeldin A / Q000494:pharmacology; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D019791:Guanidine / Q000494:pharmacology; D000882:Haplorhini; D030041:Hepatitis A virus / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology*; D007668:Kidney; D012313:RNA / Q000378:metabolism*; D012337:RNA, Ribosomal, 18S / Q000378:metabolism; D012367:RNA, Viral / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
D012337:RNA, Ribosomal, 18S; D012367:RNA, Viral; C096539:VP1 protein, hepatitis A virus; D015678:Viral Structural Proteins; D020126:Brefeldin A; D012313:RNA; D003513:Cycloheximide; D019791:Guanidine