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<title xml:lang="en">Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication</title>
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<name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name>
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<name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name>
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<name sortKey="Hidari, Kazuya I P Jwa" sort="Hidari, Kazuya I P Jwa" uniqKey="Hidari K" first="Kazuya I.-P. Jwa" last="Hidari">Kazuya I.-P. Jwa Hidari</name>
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<name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name>
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<name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
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<name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name>
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<title xml:lang="en" level="a" type="main">Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication</title>
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<name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name>
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<name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name>
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<name sortKey="Guo, Chao Tan" sort="Guo, Chao Tan" uniqKey="Guo C" first="Chao-Tan" last="Guo">Chao-Tan Guo</name>
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<name sortKey="Hidari, Kazuya I P Jwa" sort="Hidari, Kazuya I P Jwa" uniqKey="Hidari K" first="Kazuya I.-P. Jwa" last="Hidari">Kazuya I.-P. Jwa Hidari</name>
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<name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name>
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<name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
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<name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name>
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<title level="j">Journal of Virology</title>
<idno type="ISSN">0022-538X</idno>
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<p>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-
<italic>N</italic>
-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-
<italic>N</italic>
-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.</p>
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<pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front>
<journal-meta>
<journal-id journal-id-type="nlm-ta">J Virol</journal-id>
<journal-id journal-id-type="publisher-id">jvi</journal-id>
<journal-title>Journal of Virology</journal-title>
<issn pub-type="ppub">0022-538X</issn>
<issn pub-type="epub">1098-5514</issn>
<publisher>
<publisher-name>American Society for Microbiology</publisher-name>
</publisher>
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<article-id pub-id-type="pmid">16140748</article-id>
<article-id pub-id-type="pmc">1212606</article-id>
<article-id pub-id-type="publisher-id">0492-05</article-id>
<article-id pub-id-type="doi">10.1128/JVI.79.18.11705-11715.2005</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Virus-Cell Interactions</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Suzuki</surname>
<given-names>Takashi</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="fn" rid="fn1"></xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Takahashi</surname>
<given-names>Tadanobu</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="fn" rid="fn1"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Guo</surname>
<given-names>Chao-Tan</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="aff" rid="aff1">3</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Hidari</surname>
<given-names>Kazuya I.-P. Jwa</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Miyamoto</surname>
<given-names>Daisei</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Goto</surname>
<given-names>Hideo</given-names>
</name>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="aff" rid="aff1">4</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Kawaoka</surname>
<given-names>Yoshihiro</given-names>
</name>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="aff" rid="aff1">4</xref>
<xref ref-type="aff" rid="aff1">5</xref>
<xref ref-type="aff" rid="aff1">6</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Suzuki</surname>
<given-names>Yasuo</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff1">2</xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
</contrib-group>
<aff id="aff1">Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century, Shizuoka, Japan,
<label>1</label>
CREST, Japan Science and Technology Agency, Saitama, Japan,
<label>2</label>
Institute of Bioengineering, Zhejiang Academy of Medical Sciences, Hang Zhou, China,
<label>3</label>
Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo,
<label>4</label>
International Research Center for Infectious Diseases, Tokyo, Japan,
<label>5</label>
Department of Pathobiological Sciences, University of Wisconsin—Madison, Madison, Wisconsin
<label>6</label>
</aff>
<author-notes>
<fn id="cor1">
<label>*</label>
<p>Corresponding author. Mailing address: Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences, Shizuoka 422-8526, Japan. Phone: 81 54 264 5722. Fax: 81 54 264 5723. E-mail for T. Suzuki:
<email>Suzukit@u-shizuoka-ken.ac.jp</email>
.</p>
</fn>
<fn id="fn1">
<label></label>
<p>These authors contributed equally to this work.</p>
</fn>
</author-notes>
<pub-date pub-type="ppub">
<month>9</month>
<year>2005</year>
</pub-date>
<volume>79</volume>
<issue>18</issue>
<fpage>11705</fpage>
<lpage>11715</lpage>
<history>
<date date-type="received">
<day>10</day>
<month>3</month>
<year>2005</year>
</date>
<date date-type="accepted">
<day>20</day>
<month>6</month>
<year>2005</year>
</date>
</history>
<copyright-statement>Copyright © 2005, American Society for Microbiology</copyright-statement>
<copyright-year>2005</copyright-year>
<abstract>
<p>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-
<italic>N</italic>
-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-
<italic>N</italic>
-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.</p>
</abstract>
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