Interaction between polypeptide 3ABC and the 5′-terminal structural elements of the genome of Aichi virus

Implication for negative-strand RNA synthesis

Shigeo Nagashima, Jun Sasaki, Koki Taniguchi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Secondary structural elements at the 5′ end of picornavirus genomic RNA function as cis-acting replication elements and are known to interact specifically with viral P3 proteins in several picornaviruses. In poliovirus, ribonucleoprotein complex formation at the 5′ end of the genome is required for negative-strand synthesis. We have previously shown that the 5′-end 115 nucleotides of the Aichi virus genome, which are predicted to fold into two stem-loops (SL-A and SL-C) and one pseudoknot (PK-B), act as a cis-acting replication element and that correct folding of these structures is required for negative-strand synthesis. In this study, we investigated the interaction between the 5′-terminal 120 nucleotides of the genome and the P3 proteins, 3AB, 3ABC, 3C, and 3CD, by gel shift assay and Northwestern analysis. The results showed that 3ABC and 3CD bound to the 5′-terminal region specifically. The binding of 3ABC was observed on both assays, while that of 3CD was detected only on Northwestern analysis. No binding of 3AB or 3C was observed. Binding assays using mutant RNAs demonstrated that disruption of the base pairings of the stem of SL-A and one of the two stem segments of PK-B (stem-B1) abolished the 3ABC binding. In addition, the specific nucleotide sequence of stem-B1 was responsible for the efficient 3ABC binding. These results suggest that the interaction of 3ABC with the 5′-terminal region of the genome is involved in negative-strand synthesis. On the other hand, the ability of 3CD to interact with the 5′-terminal region did not correlate with the RNA replication ability.

Original languageEnglish
Pages (from-to)6161-6171
Number of pages11
JournalJournal of Virology
Volume82
Issue number13
DOIs
Publication statusPublished - 01-07-2008

Fingerprint

Aichi virus
Kobuvirus
polypeptides
Genome
RNA
Picornaviridae
Peptides
stems
synthesis
genome
Nucleotides
assays
nucleotides
Ribonucleoproteins
Poliovirus
Viral Proteins
Enterovirus C
ribonucleoproteins
Base Pairing
viral proteins

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

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abstract = "Secondary structural elements at the 5′ end of picornavirus genomic RNA function as cis-acting replication elements and are known to interact specifically with viral P3 proteins in several picornaviruses. In poliovirus, ribonucleoprotein complex formation at the 5′ end of the genome is required for negative-strand synthesis. We have previously shown that the 5′-end 115 nucleotides of the Aichi virus genome, which are predicted to fold into two stem-loops (SL-A and SL-C) and one pseudoknot (PK-B), act as a cis-acting replication element and that correct folding of these structures is required for negative-strand synthesis. In this study, we investigated the interaction between the 5′-terminal 120 nucleotides of the genome and the P3 proteins, 3AB, 3ABC, 3C, and 3CD, by gel shift assay and Northwestern analysis. The results showed that 3ABC and 3CD bound to the 5′-terminal region specifically. The binding of 3ABC was observed on both assays, while that of 3CD was detected only on Northwestern analysis. No binding of 3AB or 3C was observed. Binding assays using mutant RNAs demonstrated that disruption of the base pairings of the stem of SL-A and one of the two stem segments of PK-B (stem-B1) abolished the 3ABC binding. In addition, the specific nucleotide sequence of stem-B1 was responsible for the efficient 3ABC binding. These results suggest that the interaction of 3ABC with the 5′-terminal region of the genome is involved in negative-strand synthesis. On the other hand, the ability of 3CD to interact with the 5′-terminal region did not correlate with the RNA replication ability.",
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Interaction between polypeptide 3ABC and the 5′-terminal structural elements of the genome of Aichi virus : Implication for negative-strand RNA synthesis. / Nagashima, Shigeo; Sasaki, Jun; Taniguchi, Koki.

In: Journal of Virology, Vol. 82, No. 13, 01.07.2008, p. 6161-6171.

Research output: Contribution to journalArticle

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