Dataset Information

Role of RNA structures in genome terminal sequences of the hepatitis C virus for replication and assembly.

ABSTRACT: Hepatitis C virus (HCV) is a positive-strand RNA virus replicating its genome via a negative-strand [(-)] intermediate. Little is known about replication signals residing in the 3' end of HCV (-) RNA. Recent studies identified seven stem-loop structures (SL-I', -IIz', -IIy', -IIIa', -IIIb', -IIIcdef', and -IV') in this region. In the present study, we mapped the minimal region required for RNA replication to SL-I' and -IIz', functionally confirmed the SL-IIz' structure, and identified SL-IIIa' to -IV' as auxiliary replication elements. In addition, we show that the 5' nontranslated region of the genome most likely does not contain cis-acting RNA structures required for RNA packaging into infectious virions.

SUBMITTER: Friebe P

PROVIDER: S-EPMC2772684 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Role of RNA structures in genome terminal sequences of the hepatitis C virus for replication and assembly.

Friebe Peter P Bartenschlager Ralf R

Journal of virology 20090909 22

Hepatitis C virus (HCV) is a positive-strand RNA virus replicating its genome via a negative-strand [(-)] intermediate. Little is known about replication signals residing in the 3' end of HCV (-) RNA. Recent studies identified seven stem-loop structures (SL-I', -IIz', -IIy', -IIIa', -IIIb', -IIIcdef', and -IV') in this region. In the present study, we mapped the minimal region required for RNA replication to SL-I' and -IIz', functionally confirmed the SL-IIz' structure, and identified SL-IIIa' t ...[more]

PMID: 19740989

Similar Datasets

Project description:Influenza A viruses are responsible for seasonal epidemics, and pandemics can arise from the transmission of novel zoonotic influenza A viruses to humans1,2. Influenza A viruses contain a segmented negative-sense RNA genome, which is transcribed and replicated by the viral-RNA-dependent RNA polymerase (FluPolA) composed of PB1, PB2 and PA subunits3-5. Although the high-resolution crystal structure of FluPolA of bat influenza A virus has previously been reported6, there are no complete structures available for human and avian FluPolA. Furthermore, the molecular mechanisms of genomic viral RNA (vRNA) replication-which proceeds through a complementary RNA (cRNA) replicative intermediate, and requires oligomerization of the polymerase7-10-remain largely unknown. Here, using crystallography and cryo-electron microscopy, we determine the structures of FluPolA from human influenza A/NT/60/1968 (H3N2) and avian influenza A/duck/Fujian/01/2002 (H5N1) viruses at a resolution of 3.0-4.3 Å, in the presence or absence of a cRNA or vRNA template. In solution, FluPolA forms dimers of heterotrimers through the C-terminal domain of the PA subunit, the thumb subdomain of PB1 and the N1 subdomain of PB2. The cryo-electron microscopy structure of monomeric FluPolA bound to the cRNA template reveals a binding site for the 3' cRNA at the dimer interface. We use a combination of cell-based and in vitro assays to show that the interface of the FluPolA dimer is required for vRNA synthesis during replication of the viral genome. We also show that a nanobody (a single-domain antibody) that interferes with FluPolA dimerization inhibits the synthesis of vRNA and, consequently, inhibits virus replication in infected cells. Our study provides high-resolution structures of medically relevant FluPolA, as well as insights into the replication mechanisms of the viral RNA genome. In addition, our work identifies sites in FluPolA that could be targeted in the development of antiviral drugs.

Project description:Hepatitis C virus (HCV) preferentially replicates in the human liver and frequently causes chronic infection, often leading to cirrhosis and liver cancer. HCV is an enveloped virus classified in the genus Hepacivirus in the family Flaviviridae and has a single-stranded RNA genome of positive orientation. The HCV RNA genome is translated and replicated in the cytoplasm. Translation is controlled by the Internal Ribosome Entry Site (IRES) in the 5' untranslated region (5' UTR), while also downstream elements like the cis-replication element (CRE) in the coding region and the 3' UTR are involved in translation regulation. The cis-elements controlling replication of the viral RNA genome are located mainly in the 5'- and 3'-UTRs at the genome ends but also in the protein coding region, and in part these signals overlap with the signals controlling RNA translation. Many long-range RNA-RNA interactions (LRIs) are predicted between different regions of the HCV RNA genome, and several such LRIs are actually involved in HCV translation and replication regulation. A number of RNA cis-elements recruit cellular RNA-binding proteins that are involved in the regulation of HCV translation and replication. In addition, the liver-specific microRNA-122 (miR-122) binds to two target sites at the 5' end of the viral RNA genome as well as to at least three additional target sites in the coding region and the 3' UTR. It is involved in the regulation of HCV RNA stability, translation and replication, thereby largely contributing to the hepatotropism of HCV. However, we are still far from completely understanding all interactions that regulate HCV RNA genome translation, stability, replication and encapsidation. In particular, many conclusions on the function of cis-elements in HCV replication have been obtained using full-length HCV genomes or near-full-length replicon systems. These include both genome ends, making it difficult to decide if a cis-element in question acts on HCV replication when physically present in the plus strand genome or in the minus strand antigenome. Therefore, it may be required to use reduced systems that selectively focus on the analysis of HCV minus strand initiation and/or plus strand initiation.

Dataset Information

Role of RNA structures in genome terminal sequences of the hepatitis C virus for replication and assembly.

Publications

Role of RNA structures in genome terminal sequences of the hepatitis C virus for replication and assembly.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets