{"created":"2023-07-27T06:54:58.285001+00:00","id":50575,"links":{},"metadata":{"_buckets":{"deposit":"1a19ee2c-dcb2-4470-8c81-4ed531eae27e"},"_deposit":{"created_by":18,"id":"50575","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"50575"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00050575","sets":["2812:2813:2832"]},"author_link":["28003","91554"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2003-09-16","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"2000-2001","bibliographic_titles":[{"bibliographic_title":"平成13(2001)年度 科学研究費補助金 基盤研究(C) 研究成果報告書概要"},{"bibliographic_title":"2001 Fiscal Year Final Research Report Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"インフルエンザウイルス非粒子構成タンパク質NS1は分子量わずか26kDであるが多くの機能を持つ.その内の1つとして,我々はNS1がRNA結合活性を持つことを明らかにしたが,最近,このRNA結合能が,インフルエンザウイルスのインターフェロン抵抗性と密接に関係していることが明かにされ,このウイルスの病原性に対するNS1の重要性の認識が高まった.このように,多機能タンパク質NS1は種々の宿主細胞機能を制御することにより,ウイルス増殖をいろいろな段階で調節する.これまで,病原性決定要因として血球凝集素の切断が主に研究されて来たのに対し,NS1をターゲットとすることにより新たな局面を目指すものである.本研究ではNS1のRNA結合能を詳細に解析し,病原性の指標としてのPKR活性化との関係を調べた.\n(1)C末側によるNS1のPKR活性化阻害の調節機構について\nNS1のRNA結合必須領域はN末側の82アミノ酸に限局しており,PKR活性化を強く阻害する.このRNA結合活性は引き続く60以上のアミノ酸配列の付加により減弱され,さらにAクラスターに結合しなくなる.(1-82)NS1,完全長NS1ともUクラスターに結合することが新たに明らかになり,vRNA分節の5'末端の5-6塩基のUクラスターに強固に結合する.\n(2)リン酸化によるNS1の機能の調節\nNS1は複数のSer/Thrのリン酸化を受けるが、その部位の同定を行った。リン酸化によりRNA結合能が低下するがPKR活性化阻害に対する影響はなかった。これらのリン酸化部位をAlaやAspに置換したNS1を作成し,それぞれの部位のリン酸化の影響を調べた.また感染細胞内の核や細胞質に分布するNS1、またvRNPやリボゾームに結合しているNS1のリン酸化の度合いを感染時間を追って定量したが、存在部位による差は認められなかった。","subitem_description_type":"Abstract"},{"subitem_description":"The influenza virus nonstructural protein NS1 has been reported to have many functions. We previously demonstrated that it has an RNA binding activity, by which it inhibits the PKR activation in the infected cell. In addition, its RNA binding activity has been recently shown to participate in the resistance against interferon in influenza virus infection, underlining NS1 as a determinant for this virus pathogenicity. In this study, we analyzed the RNA binding ability of NS1 in more detail, and related it to the activation state of PKR, which is one of the indicators of this virus pathogenicity.\n(1) Effect of C-terminal region of NS1 on its inhibition of PKR activation. The region of NS1 essential for its RNA binding was the N-terminal 82 araino acid sequence, which strongly inhibits the activation of PKR. This truncated NS1 in the presence of the following more than 60.araino acid sequence, and also the full-length NS1, attenuated this activity, and lost its poly A binding activity. It was newly disclosed that NS1'longer than N-terminal 82 amino acid,sequence has a binding ability to U clusters, and bound to the 5-6 U cluster at the 5'-terminal region of Vrna.\n(2) Effect of phosphorylation of NS1 on its functions. We identified several Ser/Thr residues which are phosphorylated in the infected cells.\nThe effects of NS1 phosphorylation at each of these residues in vitro and in vivo are now investigated by constructing Ala or Asp substitution mutants of these residues. Phosphorylation of NS1 in vitro attenuated its RNA binding activity, but had little effect on its inhibition of the PKR activation. NS1 in the infected cells is distributed in the nucleus and the cytoplasm, and is associated with VRNP and ribosomes. The phosphorylation level of NS1 of these fractions were separately determined without appreciable difference among them.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:12670279, 研究期間(年度):2000-2001","subitem_description_type":"Other"},{"subitem_description":"出典：研究課題「PKR阻害蛋白質によるインフルエンザウイルス病原性の制御とその応用」課題番号12670279\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） \n（https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-12670279/126702792001kenkyu_seika_hokoku_gaiyo/）を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学医薬保健研究域医学系","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00056884","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=60027331"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=60027331","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670279/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670279/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-12670279/126702792001kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-12670279/126702792001kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2022-04-14"}],"displaytype":"detail","filename":"ME-PR-FUKUDA-R-kaken 2003-2p.pdf","filesize":[{"value":"83.0 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-FUKUDA-R-kaken 2003-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/50575/files/ME-PR-FUKUDA-R-kaken 2003-2p.pdf"},"version_id":"3b2b7403-9e97-44d3-9702-03d0d3fb8859"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"PKR阻害蛋白質によるインフルエンザウイルス病原性の制御とその応用","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"PKR阻害蛋白質によるインフルエンザウイルス病原性の制御とその応用"},{"subitem_title":"Control of influenza virus pathogenicity by pkr inhibitor protein and its application","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2832"],"pubdate":{"attribute_name":"公開日","attribute_value":"2022-04-14"},"publish_date":"2022-04-14","publish_status":"0","recid":"50575","relation_version_is_last":true,"title":["PKR阻害蛋白質によるインフルエンザウイルス病原性の制御とその応用"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T13:16:41.235941+00:00"}