3CD, but not 3C, cleaves the VP1/2A site efficiently during Aichi virus polyprotein processing through interaction with 2A

Jun Sasaki, Kumiko Ishikawa, Koki Taniguchi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Picornavirus genomes are translated into a single large polyprotein, which is processed by virus-encoded proteases into individual functional proteins. 3C of all picornaviruses is a protease, and the leader (L) and 2A proteins of some picornaviruses are also involved in polyprotein processing. Aichi virus (AiV), which is associated with acute gastroenteritis in humans, is a member of the genus Kobuvirus of the family Picornaviridae. The AiV L and 2A proteins have already been shown to exhibit no protease activity. In this study, we investigated AiV polyprotein processing by 3C and 3CD using a cell-free translation system. 3C and 3CD were capable of processing the polyprotein in trans; 3C, however, cleaved the VP1/2A site inefficiently, while 3CD cleaved this site almost completely. Mammalian two-hybrid and coimmunoprecipitation assays showed an interaction between 2A and 3CD. Using a 3CD mutant and various 2A mutants of substrate proteins, we showed a clear correlation between the 2A-3CD interaction and the VP1/2A cleavage by 3CD. Thus, this study suggests that tight interaction of 3CD with the 2A region of a precursor protein is required for efficient cleavage at the VP1/2A site.

Original languageEnglish
Pages (from-to)592-598
Number of pages7
JournalVirus Research
Volume163
Issue number2
DOIs
Publication statusPublished - 01-02-2012

Fingerprint

Kobuvirus
Picornaviridae
Polyproteins
Peptide Hydrolases
Two-Hybrid System Techniques
Proteins
Protein Precursors
Cell-Free System
Gastroenteritis
Mutant Proteins
Genome
Viruses

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases
  • Cancer Research

Cite this

@article{983ec39f51b1421ab6e871cbbcb339ed,
title = "3CD, but not 3C, cleaves the VP1/2A site efficiently during Aichi virus polyprotein processing through interaction with 2A",
abstract = "Picornavirus genomes are translated into a single large polyprotein, which is processed by virus-encoded proteases into individual functional proteins. 3C of all picornaviruses is a protease, and the leader (L) and 2A proteins of some picornaviruses are also involved in polyprotein processing. Aichi virus (AiV), which is associated with acute gastroenteritis in humans, is a member of the genus Kobuvirus of the family Picornaviridae. The AiV L and 2A proteins have already been shown to exhibit no protease activity. In this study, we investigated AiV polyprotein processing by 3C and 3CD using a cell-free translation system. 3C and 3CD were capable of processing the polyprotein in trans; 3C, however, cleaved the VP1/2A site inefficiently, while 3CD cleaved this site almost completely. Mammalian two-hybrid and coimmunoprecipitation assays showed an interaction between 2A and 3CD. Using a 3CD mutant and various 2A mutants of substrate proteins, we showed a clear correlation between the 2A-3CD interaction and the VP1/2A cleavage by 3CD. Thus, this study suggests that tight interaction of 3CD with the 2A region of a precursor protein is required for efficient cleavage at the VP1/2A site.",
author = "Jun Sasaki and Kumiko Ishikawa and Koki Taniguchi",
year = "2012",
month = "2",
day = "1",
doi = "10.1016/j.virusres.2011.12.013",
language = "English",
volume = "163",
pages = "592--598",
journal = "Virus Research",
issn = "0168-1702",
publisher = "Elsevier",
number = "2",

}

3CD, but not 3C, cleaves the VP1/2A site efficiently during Aichi virus polyprotein processing through interaction with 2A. / Sasaki, Jun; Ishikawa, Kumiko; Taniguchi, Koki.

In: Virus Research, Vol. 163, No. 2, 01.02.2012, p. 592-598.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 3CD, but not 3C, cleaves the VP1/2A site efficiently during Aichi virus polyprotein processing through interaction with 2A

AU - Sasaki, Jun

AU - Ishikawa, Kumiko

AU - Taniguchi, Koki

PY - 2012/2/1

Y1 - 2012/2/1

N2 - Picornavirus genomes are translated into a single large polyprotein, which is processed by virus-encoded proteases into individual functional proteins. 3C of all picornaviruses is a protease, and the leader (L) and 2A proteins of some picornaviruses are also involved in polyprotein processing. Aichi virus (AiV), which is associated with acute gastroenteritis in humans, is a member of the genus Kobuvirus of the family Picornaviridae. The AiV L and 2A proteins have already been shown to exhibit no protease activity. In this study, we investigated AiV polyprotein processing by 3C and 3CD using a cell-free translation system. 3C and 3CD were capable of processing the polyprotein in trans; 3C, however, cleaved the VP1/2A site inefficiently, while 3CD cleaved this site almost completely. Mammalian two-hybrid and coimmunoprecipitation assays showed an interaction between 2A and 3CD. Using a 3CD mutant and various 2A mutants of substrate proteins, we showed a clear correlation between the 2A-3CD interaction and the VP1/2A cleavage by 3CD. Thus, this study suggests that tight interaction of 3CD with the 2A region of a precursor protein is required for efficient cleavage at the VP1/2A site.

AB - Picornavirus genomes are translated into a single large polyprotein, which is processed by virus-encoded proteases into individual functional proteins. 3C of all picornaviruses is a protease, and the leader (L) and 2A proteins of some picornaviruses are also involved in polyprotein processing. Aichi virus (AiV), which is associated with acute gastroenteritis in humans, is a member of the genus Kobuvirus of the family Picornaviridae. The AiV L and 2A proteins have already been shown to exhibit no protease activity. In this study, we investigated AiV polyprotein processing by 3C and 3CD using a cell-free translation system. 3C and 3CD were capable of processing the polyprotein in trans; 3C, however, cleaved the VP1/2A site inefficiently, while 3CD cleaved this site almost completely. Mammalian two-hybrid and coimmunoprecipitation assays showed an interaction between 2A and 3CD. Using a 3CD mutant and various 2A mutants of substrate proteins, we showed a clear correlation between the 2A-3CD interaction and the VP1/2A cleavage by 3CD. Thus, this study suggests that tight interaction of 3CD with the 2A region of a precursor protein is required for efficient cleavage at the VP1/2A site.

UR - http://www.scopus.com/inward/record.url?scp=84856555220&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856555220&partnerID=8YFLogxK

U2 - 10.1016/j.virusres.2011.12.013

DO - 10.1016/j.virusres.2011.12.013

M3 - Article

C2 - 22226945

AN - SCOPUS:84856555220

VL - 163

SP - 592

EP - 598

JO - Virus Research

JF - Virus Research

SN - 0168-1702

IS - 2

ER -