{"created":"2023-07-27T06:58:38.221810+00:00","id":57407,"links":{},"metadata":{"_buckets":{"deposit":"b5da815a-07dd-4c9f-a6d8-bfebf0951e01"},"_deposit":{"created_by":18,"id":"57407","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"57407"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00057407","sets":["2812:2813:2830"]},"author_link":["95635","48959"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2005-04-18","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"2002 – 2003","bibliographic_titles":[{"bibliographic_title":"平成15(2003)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":"2003 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":"真核生物RNAポリメラーゼII(RNAP II)最大サブユニットC-末端領域(CTD)は、種々のCTDキナーゼによってリン酸化されることによって、mRNA前駆体の転写とプロセシングおよび両者のカップリングの制御に関わっている。私は、CTDリン酸化による遺伝子発現調節の分子機構にアプローチするために、リン酸化に依存してCTDに結合するヒト新規因子を同定しその機能検索を行っている。本研究は、リン酸化CTD結合因子として新規に同定したヒト核蛋白質PCIF1の生物機能を、高頻度相同組み換えを起こすことが知られているトリB細胞株DT40を用いたジーンノックアウト、および組換えタンパク質を用いた生化学的解析によって検索することを目的としており、解析の成果および中間結果を以下に報告する。\n(1)トリPCIF1遺伝子のホモノックアウトDT40細胞株、およびテトラサイクリン添加によってPCIF1の発現をオフにできるコンディショナルノックアウトDT40細胞株を樹立した。(2)PCIF1ノックアウトDT40細胞を用い、PCIF1非存在下に於ける細胞増殖能、細胞全体のRNAP IIのリン酸化とヒストンH3のメチル化の程度、および熱ショック下における熱ショックタンパク質(HSP)遺伝子の発現誘導能の変化を解析した。その結果、PCIF1は細胞の増殖に必須なタンパク質でないことが示唆され、PCIF1ノックアウトによる細胞全体のRNAP IIのリン酸化およびヒストンH3のメチル化の程度、更にHSP mRNAの発現誘導に対する影響は見出されなかった。(3)トリPCIF1の発現減少・消失に伴って、細胞周期調節に関わるペプチジルイソメラーゼPin1の発現の亢進が観察された。(4)PCIF1発現・精製のための組換えバキュロウイルスを作製した。またTAPタグを持つ外来性PCIF1を発現誘導出来るヒト安定細胞株を樹立した。","subitem_description_type":"Abstract"},{"subitem_description":"Phosphorylation of the carboxy-terminal domain (CTD) of RNA polymerase II largest subunit has important roles both in transcription and in coupling transcription to pre-mRNA processing. To better understand the molecular mechanism in which transcription coordinates with pre-mRNA processing, I have identified and characterized human factors that can directly interact with the phosphorylated CTD (pCTD).Recently I reported a novel human protein PCIF1 as a pCTD interacting factor. To investigate cellular functions of PCIF1, I disrupted the PCIF1 gene in the chicken B-cell line DT4O. I have established two independent mutant DT40 cell lines in which all three copies of PCIF1 gene are disrupted by homologous recombination. One clone is a homozygous PCIF1-null mutant and the other is a conditional knock out cell line in which PCIF1 protein is expressed from a chicken PCIF1 cDNA under control of a tetracycline-repressible promoter.Addition of doxycyclineto the cells results in depletion of PCIF1 protein within several days but not in growth defect. Thus, chicken PCIF1 is not essential for cell growth. Depletion of PC1FI in DT40 cells did not significantly affect neither the phosphorylation status of the CTD nor the methylation status of the histone H3 N-terminus region.PCIF1-deficient cells exhibited normal heat shock-response, as measured by inducible expression of heat shock genes.","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:14580682, 研究期間(年度):2002 – 2003","subitem_description_type":"Other"},{"subitem_description":"出典：「新規リン酸化RNAポリメラーゼII結合蛋白質の機能解析」研究成果報告書　課題番号14580682\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/ja/report/KAKENHI-PROJECT-14580682/145806822003kenkyu_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":"金沢大学がん研究所","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00063677","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/ja/search/?kw=00218851"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/search/?kw=00218851","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-14580682/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-14580682/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-14580682/145806822003kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-14580682/145806822003kenkyu_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":"2021-10-22"}],"displaytype":"detail","filename":"CA-PR-HIROSE-Y-kaken 2005-2p.pdf","filesize":[{"value":"77.8 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"CA-PR-HIROSE-Y-kaken 2005-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/57407/files/CA-PR-HIROSE-Y-kaken 2005-2p.pdf"},"version_id":"fd0ba30d-6c62-43cf-9965-574ec74e27a7"}]},"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":"新規リン酸化RNAポリメラーゼII結合蛋白質の機能解析","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"新規リン酸化RNAポリメラーゼII結合蛋白質の機能解析"},{"subitem_title":"Functional characterization of phosphorylated RNA polyrnerase II interacting factors","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2830"],"pubdate":{"attribute_name":"公開日","attribute_value":"2021-10-22"},"publish_date":"2021-10-22","publish_status":"0","recid":"57407","relation_version_is_last":true,"title":["新規リン酸化RNAポリメラーゼII結合蛋白質の機能解析"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:35:31.787393+00:00"}