The internal ribosome entry site (IRES) of hepatitis C virus visualized by electron microscopy

• Published in: RNA (Cambridge) 2001-05, Vol.7 (5), p.661-670, Article S1355838201001406
• Main Authors: BEALES, LUCY P., ROWLANDS, DAVID J., HOLZENBURG, ANDREAS
• Format: Article
• Language: English
• Subjects: 5' Untranslated Regions - chemistry; 5' Untranslated Regions - ultrastructure; Aphthovirus - genetics; Base Sequence; Hepacivirus - genetics; Hepatitis C virus; Humans; Microscopy, Electron - methods; Molecular Sequence Data; Nucleic Acid Conformation; pyrimidine tract-binding protein; Ribosomes - metabolism; RNA, Viral - chemistry; RNA, Viral - ultrastructure
• ISSN: 1355-8382; 1469-9001
• DOI: 10.1017/S1355838201001406

Translation of hepatitis C virus (HCV) RNA is initiated via the internal ribosome entry site (IRES), located within the 5′ untranslated region. Although the secondary structure of this element has been predicted, little information on the tertiary structure is available. Here we report the first structural characterization of the HCV IRES using electron microscopy. In vitro transcribed RNA appeared as particles with characteristic morphology and gold labeling using a specific oligonucleotide confirmed them to be HCV IRES. Dimerization of the IRES by hybridization with tandem repeat oligonucleotides allowed the identification of domain III and an assignment of domains II and IV to distinct regions within the molecule. Using immunogold labeling, the pyrimidine tract binding protein (PTB) was shown to bind to domain III. Structure–function relationships based on the flexible hinge between domains II and III are suggested. Finally, the architecture of the HCV IRES was seen to be markedly different from that of a picornavirus, foot-and-mouth disease virus (FMDV).