Sialidase activity of influenza A virus in an endocytic pathway enhances viral replication
Identifieur interne : 000024 ( PascalFrancis/Curation ); précédent : 000023; suivant : 000025Sialidase activity of influenza A virus in an endocytic pathway enhances viral replication
Auteurs : Takashi Suzuki [Japon] ; Tadanobu Takahashi [Japon] ; Chao-Tan Guo [Japon, République populaire de Chine] ; Kazuya I.-P. Jwa Hidari [Japon] ; Daisei Miyamoto [Japon] ; Hideo Goto [Japon] ; Yoshihiro Kawaoka [Japon, États-Unis] ; Yasuo Suzuki [Japon]Source :
- Journal of virology [ 0022-538X ] ; 2005.
Descripteurs français
- Pascal (Inist)
English descriptors
Abstract
N2 neuraminidase (NA) genes of the 1957 and 1968 pandemic influenza virus strains possessed avian-like low-pH stability of sialidase activity, unlike most epidemic strains. We generated four reverse-genetics viruses from a genetic background of A/WSN/33 (H1N1) that included parental N2 NAs of 1968 pandemic (H3N2) and epidemic (H2N2) strains or their counterpart N2 NAs in which the low-pH stability of the sialidase activity was changed by substitutions of one or two amino acid residues. We found that the transfectant viruses bearing low-pH-stable sialidase (WSN/Stable-NAs) showed 25- to 80-times-greater ability to replicate in Madin-Darby canine kidney (MDCK) cells than did the transfectant viruses bearing low-pH-unstable sialidase (WSN/ Unstable-NAs). Enzymatic activities of WSN/Stable-NAs were detected in endosomes of MDCK cells after 90 min of virus internalization by in situ fluorescent detection with 5-bromo-4-chloro-indole-3-yl-α-N-acetylneuraminic acid and Fast Red Violet LB. Inhibition of sialidase activity of WSN/Stable-NAs on the endocytic pathway by pretreatment with 4-guanidino-2,4-dideoxy-N-acetylneuraminic acid (zanamivir) resulted in a significant decrease in progeny viruses. In contrast, the enzymatic activities of WSN/Unstable-NAs, the replication of which had no effect on pretreatment with zanamivir, were undetectable in cells under the same conditions. Hemadsorption assays of transfectant-virus-infected cells revealed that the low-pH stability of the sialidase had no effect on the process of removal of sialic acid from hemagglutinin in the Golgi regions. Moreover, high titers of viruses were recovered from the lungs of mice infected with WSN/Stable-NAs on day 3 after intranasal inoculation, but WSN/Unstable-NAs were cleared from the lungs of the mice. These results indicate that sialidase activity in late endosome/lysosome traffic enhances influenza A virus replication.
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Sialidase activity of influenza A virus in an endocytic pathway enhances viral replication</title><author><name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Guo, Chao Tan" sort="Guo, Chao Tan" uniqKey="Guo C" first="Chao-Tan" last="Guo">Chao-Tan Guo</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Institute of Bioengineering, Zhejiang Academy of Medical Sciences</s1><s2>Hang Zhou</s2><s3>CHN</s3><sZ>3 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Jwa Hidari, Kazuya I P" sort="Jwa Hidari, Kazuya I P" uniqKey="Jwa Hidari K" first="Kazuya I.-P." last="Jwa Hidari">Kazuya I.-P. Jwa Hidari</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Miyamoto, Daisei" sort="Miyamoto, Daisei" uniqKey="Miyamoto D" first="Daisei" last="Miyamoto">Daisei Miyamoto</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>International Research Center for Infectious Diseases</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Department of Pathobiological Sciences, University of Wisconsin-Madison</s1><s2>Madison, Wisconsin</s2><s3>USA</s3><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country></affiliation></author><author><name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">05-0406988</idno><date when="2005">2005</date><idno type="stanalyst">PASCAL 05-0406988 INIST</idno><idno type="RBID">Pascal:05-0406988</idno><idno type="wicri:Area/PascalFrancis/Corpus">000050</idno><idno type="wicri:Area/PascalFrancis/Curation">000024</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Sialidase activity of influenza A virus in an endocytic pathway enhances viral replication</title><author><name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Guo, Chao Tan" sort="Guo, Chao Tan" uniqKey="Guo C" first="Chao-Tan" last="Guo">Chao-Tan Guo</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Institute of Bioengineering, Zhejiang Academy of Medical Sciences</s1><s2>Hang Zhou</s2><s3>CHN</s3><sZ>3 aut.</sZ></inist:fA14><country>République populaire de Chine</country></affiliation></author><author><name sortKey="Jwa Hidari, Kazuya I P" sort="Jwa Hidari, Kazuya I P" uniqKey="Jwa Hidari K" first="Kazuya I.-P." last="Jwa Hidari">Kazuya I.-P. Jwa Hidari</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Miyamoto, Daisei" sort="Miyamoto, Daisei" uniqKey="Miyamoto D" first="Daisei" last="Miyamoto">Daisei Miyamoto</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author><author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>International Research Center for Infectious Diseases</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>7 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Department of Pathobiological Sciences, University of Wisconsin-Madison</s1><s2>Madison, Wisconsin</s2><s3>USA</s3><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country></affiliation></author><author><name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country></affiliation></author></analytic><series><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Endocytosis</term><term>Exo-α-sialidase</term><term>Influenza A virus</term><term>Microbiology</term><term>Replication</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Virus grippal A</term><term>Exo-α-sialidase</term><term>Endocytose</term><term>Réplication</term><term>Microbiologie</term><term>Virologie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">N2 neuraminidase (NA) genes of the 1957 and 1968 pandemic influenza virus strains possessed avian-like low-pH stability of sialidase activity, unlike most epidemic strains. We generated four reverse-genetics viruses from a genetic background of A/WSN/33 (H1N1) that included parental N2 NAs of 1968 pandemic (H3N2) and epidemic (H2N2) strains or their counterpart N2 NAs in which the low-pH stability of the sialidase activity was changed by substitutions of one or two amino acid residues. We found that the transfectant viruses bearing low-pH-stable sialidase (WSN/Stable-NAs) showed 25- to 80-times-greater ability to replicate in Madin-Darby canine kidney (MDCK) cells than did the transfectant viruses bearing low-pH-unstable sialidase (WSN/ Unstable-NAs). Enzymatic activities of WSN/Stable-NAs were detected in endosomes of MDCK cells after 90 min of virus internalization by in situ fluorescent detection with 5-bromo-4-chloro-indole-3-yl-α-N-acetylneuraminic acid and Fast Red Violet LB. Inhibition of sialidase activity of WSN/Stable-NAs on the endocytic pathway by pretreatment with 4-guanidino-2,4-dideoxy-N-acetylneuraminic acid (zanamivir) resulted in a significant decrease in progeny viruses. In contrast, the enzymatic activities of WSN/Unstable-NAs, the replication of which had no effect on pretreatment with zanamivir, were undetectable in cells under the same conditions. Hemadsorption assays of transfectant-virus-infected cells revealed that the low-pH stability of the sialidase had no effect on the process of removal of sialic acid from hemagglutinin in the Golgi regions. Moreover, high titers of viruses were recovered from the lungs of mice infected with WSN/Stable-NAs on day 3 after intranasal inoculation, but WSN/Unstable-NAs were cleared from the lungs of the mice. These results indicate that sialidase activity in late endosome/lysosome traffic enhances influenza A virus replication.</div></front></TEI><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0022-538X</s0></fA01><fA03 i2="1"><s0>J. virol.</s0></fA03><fA05><s2>79</s2></fA05><fA06><s2>18</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Sialidase activity of influenza A virus in an endocytic pathway enhances viral replication</s1></fA08><fA11 i1="01" i2="1"><s1>SUZUKI (Takashi)</s1></fA11><fA11 i1="02" i2="1"><s1>TAKAHASHI (Tadanobu)</s1></fA11><fA11 i1="03" i2="1"><s1>GUO (Chao-Tan)</s1></fA11><fA11 i1="04" i2="1"><s1>JWA HIDARI (Kazuya I.-P.)</s1></fA11><fA11 i1="05" i2="1"><s1>MIYAMOTO (Daisei)</s1></fA11><fA11 i1="06" i2="1"><s1>GOTO (Hideo)</s1></fA11><fA11 i1="07" i2="1"><s1>KAWAOKA (Yoshihiro)</s1></fA11><fA11 i1="08" i2="1"><s1>SUZUKI (Yasuo)</s1></fA11><fA14 i1="01"><s1>Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century</s1><s2>Shizuoka</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></fA14><fA14 i1="02"><s1>CREST, Japan Science and Technology Agency</s1><s2>Saitama</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></fA14><fA14 i1="03"><s1>Institute of Bioengineering, Zhejiang Academy of Medical Sciences</s1><s2>Hang Zhou</s2><s3>CHN</s3><sZ>3 aut.</sZ></fA14><fA14 i1="04"><s1>Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ></fA14><fA14 i1="05"><s1>International Research Center for Infectious Diseases</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>7 aut.</sZ></fA14><fA14 i1="06"><s1>Department of Pathobiological Sciences, University of Wisconsin-Madison</s1><s2>Madison, Wisconsin</s2><s3>USA</s3><sZ>7 aut.</sZ></fA14><fA20><s1>11705-11715</s1></fA20><fA21><s1>2005</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>13592</s2><s5>354000131591580170</s5></fA43><fA44><s0>0000</s0><s1>© 2005 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>34 ref.</s0></fA45><fA47 i1="01" i2="1"><s0>05-0406988</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Journal of virology</s0></fA64><fA66 i1="01"><s0>USA</s0></fA66><fC01 i1="01" l="ENG"><s0>N2 neuraminidase (NA) genes of the 1957 and 1968 pandemic influenza virus strains possessed avian-like low-pH stability of sialidase activity, unlike most epidemic strains. We generated four reverse-genetics viruses from a genetic background of A/WSN/33 (H1N1) that included parental N2 NAs of 1968 pandemic (H3N2) and epidemic (H2N2) strains or their counterpart N2 NAs in which the low-pH stability of the sialidase activity was changed by substitutions of one or two amino acid residues. We found that the transfectant viruses bearing low-pH-stable sialidase (WSN/Stable-NAs) showed 25- to 80-times-greater ability to replicate in Madin-Darby canine kidney (MDCK) cells than did the transfectant viruses bearing low-pH-unstable sialidase (WSN/ Unstable-NAs). Enzymatic activities of WSN/Stable-NAs were detected in endosomes of MDCK cells after 90 min of virus internalization by in situ fluorescent detection with 5-bromo-4-chloro-indole-3-yl-α-N-acetylneuraminic acid and Fast Red Violet LB. Inhibition of sialidase activity of WSN/Stable-NAs on the endocytic pathway by pretreatment with 4-guanidino-2,4-dideoxy-N-acetylneuraminic acid (zanamivir) resulted in a significant decrease in progeny viruses. In contrast, the enzymatic activities of WSN/Unstable-NAs, the replication of which had no effect on pretreatment with zanamivir, were undetectable in cells under the same conditions. Hemadsorption assays of transfectant-virus-infected cells revealed that the low-pH stability of the sialidase had no effect on the process of removal of sialic acid from hemagglutinin in the Golgi regions. Moreover, high titers of viruses were recovered from the lungs of mice infected with WSN/Stable-NAs on day 3 after intranasal inoculation, but WSN/Unstable-NAs were cleared from the lungs of the mice. These results indicate that sialidase activity in late endosome/lysosome traffic enhances influenza A virus replication.</s0></fC01><fC02 i1="01" i2="X"><s0>002A05C10</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Virus grippal A</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Influenza A virus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Influenza A virus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Exo-α-sialidase</s0><s2>FE</s2><s5>05</s5><s6>Exo-«α»-sialidase</s6></fC03><fC03 i1="02" i2="X" l="ENG"><s0>Exo-α-sialidase</s0><s2>FE</s2><s5>05</s5><s6>Exo-«α»-sialidase</s6></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Exo-α-sialidase</s0><s2>FE</s2><s5>05</s5><s6>Exo-«α»-sialidase</s6></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Endocytose</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Endocytosis</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Endocitosis</s0><s5>06</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Réplication</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Replication</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Replicación</s0><s5>07</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Microbiologie</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Microbiology</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Microbiología</s0><s5>08</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Virologie</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Virology</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Virología</s0><s5>09</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Influenzavirus A</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Influenzavirus A</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Influenzavirus A</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Orthomyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Orthomyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Orthomyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>O-Glycosidases</s0><s2>FE</s2><s6>«O»-Glycosidases</s6></fC07><fC07 i1="04" i2="X" l="ENG"><s0>O-Glycosidases</s0><s2>FE</s2><s6>«O»-Glycosidases</s6></fC07><fC07 i1="04" i2="X" l="SPA"><s0>O-Glycosidases</s0><s2>FE</s2><s6>«O»-Glycosidases</s6></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Glycosidases</s0><s2>FE</s2></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Glycosidases</s0><s2>FE</s2></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Glycosidases</s0><s2>FE</s2></fC07><fC07 i1="06" i2="X" l="FRE"><s0>Hydrolases</s0><s2>FE</s2></fC07><fC07 i1="06" i2="X" l="ENG"><s0>Hydrolases</s0><s2>FE</s2></fC07><fC07 i1="06" i2="X" l="SPA"><s0>Hydrolases</s0><s2>FE</s2></fC07><fC07 i1="07" i2="X" l="FRE"><s0>Enzyme</s0><s2>FE</s2></fC07><fC07 i1="07" i2="X" l="ENG"><s0>Enzyme</s0><s2>FE</s2></fC07><fC07 i1="07" i2="X" l="SPA"><s0>Enzima</s0><s2>FE</s2></fC07><fN21><s1>283</s1></fN21><fN44 i1="01"><s1>OTO</s1></fN44><fN82><s1>OTO</s1></fN82></pA></standard></inist></record>Pour manipuler ce document sous Unix (Dilib)
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