Modification of the trypsin cleavage site of rotavirus VP4 to a furin-sensitive form does not enhance replication efficiency

Satoshi Komoto, Mitsutaka Wakuda, Tomihiko Ide, Gen Niimi, Yoshimasa Maeno, Kyoko Higo-Moriguchi, Koki Taniguchi

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The infectivity of rotavirus (RV) is dependent on an activation process triggered by the proteolytic cleavage of its spike protein VP4. This activation cleavage is performed by exogenous trypsin in the lumen of the intestines in vivo. Here, we report the generation and characterization of a recombinant RV expressing cDNA-derived VP4 with a modified cleavage site (arginine at position 247) recognized by endogenous furin as well as exogenous trypsin. Unexpectedly, the mutant virus (KU//rVP4-R247Furin) was incapable of plaque formation without an exogenous protease, although the mutant VP4s on virions were efficiently cleaved by endogenous furin. Furthermore, KU//rVP4-R247Furin showed impaired infectivity in MA104 and CV-1 cells even in the presence of trypsin compared with the parental virus carrying authentic VP4 (KU//rVP4). Although the total titre of KU//rVP4-R247Furin was comparable to that of KU//rVP4, the extracellular titre of KU// rVP4-R247Furin was markedly lower than its cell-associated titre in comparison with that of KU// rVP4. In contrast, the two viruses showed similar growth in a furin-defective LoVo cell line. These results suggest that intracellular cleavage of VP4 by furin may be disadvantageous for RV infectivity, possibly due to an inefficient virus release process

Original languageEnglish
Pages (from-to)2914-2921
Number of pages8
JournalJournal of General Virology
Volume92
Issue number12
DOIs
Publication statusPublished - 01-12-2011

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Furin
Rotavirus
Trypsin
Viruses
Virus Release
Virion
Intestines
Arginine
Peptide Hydrolases
Complementary DNA
Cell Line
Growth
Proteins

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

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abstract = "The infectivity of rotavirus (RV) is dependent on an activation process triggered by the proteolytic cleavage of its spike protein VP4. This activation cleavage is performed by exogenous trypsin in the lumen of the intestines in vivo. Here, we report the generation and characterization of a recombinant RV expressing cDNA-derived VP4 with a modified cleavage site (arginine at position 247) recognized by endogenous furin as well as exogenous trypsin. Unexpectedly, the mutant virus (KU//rVP4-R247Furin) was incapable of plaque formation without an exogenous protease, although the mutant VP4s on virions were efficiently cleaved by endogenous furin. Furthermore, KU//rVP4-R247Furin showed impaired infectivity in MA104 and CV-1 cells even in the presence of trypsin compared with the parental virus carrying authentic VP4 (KU//rVP4). Although the total titre of KU//rVP4-R247Furin was comparable to that of KU//rVP4, the extracellular titre of KU// rVP4-R247Furin was markedly lower than its cell-associated titre in comparison with that of KU// rVP4. In contrast, the two viruses showed similar growth in a furin-defective LoVo cell line. These results suggest that intracellular cleavage of VP4 by furin may be disadvantageous for RV infectivity, possibly due to an inefficient virus release process",
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Modification of the trypsin cleavage site of rotavirus VP4 to a furin-sensitive form does not enhance replication efficiency. / Komoto, Satoshi; Wakuda, Mitsutaka; Ide, Tomihiko; Niimi, Gen; Maeno, Yoshimasa; Higo-Moriguchi, Kyoko; Taniguchi, Koki.

In: Journal of General Virology, Vol. 92, No. 12, 01.12.2011, p. 2914-2921.

Research output: Contribution to journalArticle

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AU - Komoto, Satoshi

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