{"created":"2023-05-15T09:59:08.808714+00:00","id":3230,"links":{},"metadata":{"_buckets":{"deposit":"d59eb1bf-afa5-45b0-a36f-3831784a9add"},"_deposit":{"created_by":5,"id":"3230","owner":"5","owners":[5],"pid":{"revision_id":0,"type":"depid","value":"3230"},"status":"published"},"_oai":{"id":"oai:miyazaki-u.repo.nii.ac.jp:00003230","sets":["74","74:46","74:46:349","74:46:349:353"]},"author_link":["3752","16946","3111","16949"],"item_10002_alternative_title_1":{"attribute_name":"その他（別言語等）のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"ビワ ガンシュビョウ サイキン ノ ビョウゲンセイ イデンシ psvA ニ コード サレテイル タンパクシツ ノ キノウ カイセキ"}]},"item_10002_biblio_info_7":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2011-02-28","bibliographicIssueDateType":"Issued"},"bibliographicPageEnd":"10","bibliographicPageStart":"1","bibliographicVolumeNumber":"57","bibliographic_titles":[{"bibliographic_title":"宮崎大学農学部研究報告","bibliographic_titleLang":"ja"},{"bibliographic_title":"Bulletin of the Faculty of Agriculture University of Miyazaki","bibliographic_titleLang":"en"}]}]},"item_10002_description_5":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"The Pseudomonas syringae pv. eriobotryae (Pse) virulence gene psvA, (2193 bp), has been isolated but not been functionally characterized. The psvA gene was divided into two parts; the N-terninal region (psvAN, nucleotides (nt) 1-1386), and the C-terminal region (psvAC, nt 1387-2193). Functional analysis of the proteins encoded by psvAN and psvAC was carried out. The PsvAC shows sequence similarity to the Ulp1 endopeptidase family, which includes small ubiquitin-like modifier (SUMO) proteases. A glutathione S-transferase-PsvAC (GST-PsvAC) fusion protein cleaved tomato SUMO (T-SUMO) in vitro, while GST alone did not. The enzymatic activity of GST-PsvAC was inhibited by the cysteine protease inhibitor N-ethylmaleimide, but not by the serine protease inhibitor phenylmethanesulfonyl fluoride. Moreover, substitution of cysteine with alanine at the predicted active site abolished PsvAC enzymatic activity. These results indicate that PsvAC acts as SUMO cysteine protease. PsvAN exhibited partial homology to effector proteins. Using a adenylate cyclase (Cya) reporter system, we showed that PsvA is translocated into plant cells. To confirm our previous report that PsvA is associated with the outer membrane, a psvAN-chloramphenicol acetyltransferase (CAT) fusion gene (psvAN-CAT) was constructed and transformed into Pse PEO. The PsvAN-CAT fusion protein was detected in the outer membrane and cytoplasmic fractions by Western blot analysis using an anti-CAT antibody, whereas CAT was detected only in the cytoplasmic fraction. Furthermore, an N-terminal deletion mutant of PsvAN-CAT was not detected in the outer membrane fraction. These results suggested that PsvAN might exert its function as an outer membrane protein.","subitem_description_language":"en","subitem_description_type":"Abstract"},{"subitem_description":"ビワがんしゅ病細菌(Pse)の病原性遺伝子psvAが分離されたが,その機能は明らかにされていない. 本論文ではpsvAをN末端領域(psvAN:1-1386塩基)とC末端領域(psvAC:1387-2193塩基)に分け,それぞれの領域にコードされている蛋白質の機能解析を行った. PsvACは低分子ユビキチン様修飾因子(SUMO)プロテアーゼを含むUlp1エンドペプチダーゼファミリーと相同性を示していた. glutathione S-transferase(GST)とPsvACの融合蛋白質(GST-PsvAC)はトマトSUMOを切断したが, GSTのみでは切断されなかった. GST-PsvACの酵素活性はセリンプロテアーゼ阻害剤のphenylmethanesulfonyl fluorideでは阻害されず, システインプロテアーゼ阻害剤のN-ethylmaleimideで阻害された. 更に予想される活性部位のシステインをアラニンに置換するとPsvACの酵素活性が失われた. これらの実験結果はPsvACがSUMOシステインプロテアーゼとして機能することを示していた. また,PsvANはエフェクター蛋白質と部分的に相同生があり,アデニル酸シクラーゼ(Cya)レポーターシステムを用いてPsvAが植物細胞内に分泌されることを明らかにした. PsvAが細菌の外膜に関連していることを以前に報告したが, そのことを確かめるため, psvAN遺伝子とChloramphenicol acetyltransferase(CAT)遺伝子を融合させ,Pseに導入し, CAT抗血清を用いてウェスターンブロッティングで調べた結果, PsvAN-CAT融合蛋白質は外膜と細胞質画分に検出された.なお,CATのみの場合は細胞質画分にのみ検出された. 更にN末端側が欠落したPsvAN-CATは外膜には検出されなかった. これらの実験結果はPsvANが外膜局在性の役割を果たしていることを示唆していた.","subitem_description_language":"ja","subitem_description_type":"Abstract"}]},"item_10002_publisher_8":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"宮崎大学農学部","subitem_publisher_language":"ja"},{"subitem_publisher":"Faculty of Agriculture, University of Miyazaki","subitem_publisher_language":"en"}]},"item_10002_source_id_11":{"attribute_name":"書誌レコードID","attribute_value_mlt":[{"subitem_source_identifier":"AN00236503","subitem_source_identifier_type":"NCID"}]},"item_10002_source_id_9":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"05446066","subitem_source_identifier_type":"ISSN"}]},"item_10002_version_type_20":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"上運天, 博","creatorNameLang":"ja"},{"creatorName":"カミウンテン, ヒロシ","creatorNameLang":"ja-Kana"},{"creatorName":"Kamiunten, Hiroshi","creatorNameLang":"en"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"坂巻, 幾子","creatorNameLang":"ja"},{"creatorName":"サカマキ, イクコ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"松尾, 光弘","creatorNameLang":"ja"},{"creatorName":"マツオ, ミツヒロ","creatorNameLang":"ja-Kana"},{"creatorName":"Matsuo, Mitsuhiro","creatorNameLang":"en"}],"nameIdentifiers":[{},{}]},{"creatorNames":[{"creatorName":"Sakamaki, Ikuko","creatorNameLang":"en"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-06-21"}],"displaytype":"detail","filename":"57_1-pp.1-10.pdf","filesize":[{"value":"784.6 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"57_1-pp.1-10.pdf","url":"https://miyazaki-u.repo.nii.ac.jp/record/3230/files/57_1-pp.1-10.pdf"},"version_id":"afd4e43e-7555-4f26-a8b8-e3b433238242"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Pseudomonas syringae pv. eriobotryae, Virulence gene psvA, SUMO protease, Subcellular localization","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"ビワがんしゅ病細菌, 病原性遺伝子psvA, SUMOプロテアーゼ, 細胞内局在性","subitem_subject_language":"ja","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"Functional analysis of the protein encoded by the virulence gene psvA of Pseudomonas syringae pv. eriobotryae","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Functional analysis of the protein encoded by the virulence gene psvA of Pseudomonas syringae pv. eriobotryae","subitem_title_language":"en"},{"subitem_title":"ビワがんしゅ病細菌の病原性遺伝子psvAにコードされている蛋白質の機能解析","subitem_title_language":"ja"}]},"item_type_id":"10002","owner":"5","path":["74","46","349","353"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2012-03-16"},"publish_date":"2012-03-16","publish_status":"0","recid":"3230","relation_version_is_last":true,"title":["Functional analysis of the protein encoded by the virulence gene psvA of Pseudomonas syringae pv. eriobotryae"],"weko_creator_id":"5","weko_shared_id":2},"updated":"2023-07-30T22:53:55.844002+00:00"}