Mutational Analysis of the Bovine Hepacivirus Internal Ribosome Entry Site.
Abstract: In recent years, hepatitis C virus (HCV)-related viruses were identified in several species, including dogs, horses, bats, and rodents. In addition, a novel virus of the genus has been discovered in bovine samples and was termed bovine hepacivirus (BovHepV). Prediction of the BovHepV internal ribosome entry site (IRES) structure revealed strong similarities to the HCV IRES structure comprising domains II, IIIabcde, pseudoknot IIIf, and IV with the initiation codon AUG. Unlike HCV, only one microRNA-122 (miR-122) binding site could be identified in the BovHepV 5' nontranslated region. In this study, we analyzed the necessity of BovHepV IRES domains to initiate translation and investigated possible interactions between the IRES and core coding sequences by using a dual luciferase reporter assay. Our results suggest that such long-range interactions within the viral genome can affect IRES-driven translation. Moreover, the significance of a possible miR-122 binding to the BovHepV IRES was investigated. When analyzing translation in human Huh-7 cells with large amounts of endogenous miR-122, introduction of point mutations to the miR-122 binding site resulted in reduced translation efficiency. Similar results were observed in HeLa cells after substitution of miR-122. Nevertheless, the absence of pronounced effects in a bovine hepatocyte cell line expressing hardly any miR-122 as well suggests additional functions of this host factor in virus replication. Several members of the family , including HCV, have adapted cap-independent translation strategies to overcome canonical eukaryotic translation pathways and use -acting RNA-elements, designated viral internal ribosome entry sites (IRES), to initiate translation. Although novel hepaciviruses have been identified in different animal species, only limited information is available on their biology on molecular level. Therefore, our aim was a fundamental analysis of BovHepV IRES functions. The findings which show that functional IRES elements are also crucial for BovHepV translation expand our knowledge on molecular mechanism of hepacivirus propagation. We also studied the possible effects of one major host factor implicated in HCV pathogenesis, miR-122. The results of mutational analyses suggested that miR-122 enhances virus translation mediated by BovHepV IRES.
Copyright © 2018 American Society for Microbiology.
Publication Date: 2018-07-17 PubMed ID: 29769341PubMed Central: PMC6052288DOI: 10.1128/JVI.01974-17Google Scholar: Lookup
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Summary
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The study delves into the translation initiation and potential interactions of the Bovine Hepacivirus (BovHepV) by analyzing the functionality of its Internal Ribosome Entry Sites (IRES) and the possible impact of microRNA-122 (miR-122). The data reveal that the elements and structures of BovHepV IRES are essential for its translation, and miR-122 could be enhancing this virus translation process.
Analysis of BovHepV IRES Domains
- The researchers were mainly interested in the bovine hepatitis virus (BovHepV), which displays a similar IRES structure to the hepatitis C virus (HCV) including domains II, IIIabcde, pseudoknot IIIf, and IV with an initiation codon AUG.
- The researchers analyzed the necessity of the BovHepV IRES domains for initializing translation through a dual luciferase reporter assay.
- Results indicated potential interaction between the IRES and the virus’s core coding sequences. These internal interactions within the viral genome can affect the IRES-triggered translation.
Investigating the Role of miR-122
- A key difference in the genome of BovHepV, when compared to HCV, is that only one miR-122 binding site could be identified in BovHepV’s 5′ nontranslated region.
- The role of miR-122 was scrutinized by introducing point mutations to the miR-122 binding site and monitoring translation efficiency within human Huh-7 cells, known for their high quantity of endogenous miR-122. The mutations lead to reduced translation efficiency.
- Similar effects were seen in HeLa cells after substituting miR-122. However, due to the negligible effects in a bovine hepatocyte cell line with minimal miR-122, it suggests that this host factor might have additional roles in virus replication.
Implications for Understanding Hepacivirus Propagation
- The study expands understanding of the molecular mechanics of hepacivirus propagation by showing the importance of functional IRES elements for BovHepV and possibly other novel hepaciviruses identified in various animal species.
- The role of host factor miR-122, commonly implicated in HCV pathogenesis, was also investigated, suggesting miR-122 may enhance virus translation mediated by BovHepV IRES.
- These findings can aid in the development of treatments for various species afflicted by members of the Hepacivirus genus, including HCV in humans.
Cite This Article
APA
Baron AL, Schoeniger A, Becher P, Baechlein C.
(2018).
Mutational Analysis of the Bovine Hepacivirus Internal Ribosome Entry Site.
J Virol, 92(15), e01974-17.
https://doi.org/10.1128/JVI.01974-17 Publication
Researcher Affiliations
- Institute of Virology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Hannover, Germany.
- Institute of Biochemistry, Faculty of Veterinary Medicine, University of Leipzig, Leipzig, Germany.
- Institute of Virology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Hannover, Germany paul.becher@tiho-hannover.de christine.baechlein@tiho-hannover.de.
- German Center for Infection Research, Partner-Site Hannover-Braunschweig, Hannover, Germany.
- Institute of Virology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Hannover, Germany paul.becher@tiho-hannover.de christine.baechlein@tiho-hannover.de.
- German Center for Infection Research, Partner-Site Hannover-Braunschweig, Hannover, Germany.
MeSH Terms
- 5' Untranslated Regions
- Animals
- Cattle
- Cattle Diseases / genetics
- Cattle Diseases / metabolism
- Cattle Diseases / virology
- HeLa Cells
- Hepacivirus / genetics
- Hepacivirus / metabolism
- Humans
- Internal Ribosome Entry Sites
- RNA, Viral / genetics
- RNA, Viral / metabolism
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Citations
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