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Journal of Virology, July 2008, p. 6161-6171, Vol. 82, No. 13
0022-538X/08/$08.00+0     doi:10.1128/JVI.02151-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Interaction between Polypeptide 3ABC and the 5'-Terminal Structural Elements of the Genome of Aichi Virus: Implication for Negative-Strand RNA Synthesis{triangledown}

Shigeo Nagashima,{dagger} Jun Sasaki,* and Koki Taniguchi

Department of Virology and Parasitology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan

Received 1 October 2007/ Accepted 23 April 2008

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.

* Corresponding author. Mailing address: Department of Virology and Parasitology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan. Phone: 81-562-93-2486. Fax: 81-562-93-4008. E-mail: jsasaki{at}fujita-hu.ac.jp

{triangledown} Published ahead of print on 30 April 2008.

{dagger} Present address: Department of Hygiene, Sapporo Medical University School of Medicine, Sapporo, Hokkaido 060-8556, Japan.

Journal of Virology, July 2008, p. 6161-6171, Vol. 82, No. 13
0022-538X/08/$08.00+0     doi:10.1128/JVI.02151-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sasaki, J., Taniguchi, K. (2008). Aichi Virus 2A Protein Is Involved in Viral RNA Replication. J. Virol. 82: 9765-9769 [Abstract] [Full Text]  


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