An internal polypyrimidine-tract-binding protein-binding site in the hepatitis C virus RNA attenuates translation, which is relieved by the 3 '-untranslated sequence
T. Ito et Mmc. Lai, An internal polypyrimidine-tract-binding protein-binding site in the hepatitis C virus RNA attenuates translation, which is relieved by the 3 '-untranslated sequence, VIROLOGY, 254(2), 1999, pp. 288-296
Hepatitis C virus (HCV) RNA binds to several cellular proteins, which may r
egulate translation or replication of viral RNA. One of these is polypyrimi
dine tract-binding protein (PTB), which binds to the 5'-untranslated region
(UTR) and the 3'-end 98 nucleotides (nt) (X region) of HCV RNA. Both of th
ese PTB-binding sites regulate HCV translation. In this study, we further i
nvestigated the nature of PTB binding on HCV RNA. UV cross-linking studies
using HeLa cell extracts and a recombinant PTB showed that the PTB-5'-UTR b
inding was much weaker than the PTB-3'-UTR binding, Unexpectedly, we found
an even stronger PTB-binding site in the core-protein-coding region of HCV
RNA. The binding domain was mapped to the 3'-end of this region, which cont
ains a pyrimidine-rich sequence highly conserved among HCV isolates. Using
a set of synthetic HCV RNAs with or without this sequence in in vitro trans
lation studies, we showed that the PTB-binding sequence in the core-coding
region strongly inhibited translation of HCV RNA. This inhibition was relie
ved by the presence of the X region at the 3'-end. Furthermore, the previou
sly reported translational enhancement by the HCV 3'-UTR was more pronounce
d when this PTB-binding site was present in the RNA. These results suggest
that PTB binding to an internal site of HCV RNA provides another mechanism
for regulation of HCV translation, (C) 1999 Academic Press.