{"created":"2023-07-27T06:55:01.272842+00:00","id":50650,"links":{},"metadata":{"_buckets":{"deposit":"ccf9dde0-fad4-4131-89d8-555dd8c1353d"},"_deposit":{"created_by":18,"id":"50650","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"50650"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00050650","sets":["2812:2813:2835"]},"author_link":["25440","74433"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"1999-12-07","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"3p.","bibliographicVolumeNumber":"1997-1998","bibliographic_titles":[{"bibliographic_title":"平成10(1998)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":"1998 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":"1. 葉酸受容体(FR)遺伝子調節領域特異的結合因子の同定:(1)FR高発現細胞株であるKB細胞からの核抽出蛋白を用いて,ゲルシフト法によりFR種遺伝子Exonl上流-352/-461領域に特異的に結合する核内蛋白の存在を明らかにした.(2)さらにこの領域をオーバーラップを含む5つの断片(各36bp)に細分化したプローブを作成し,核蛋白の結合性を検索した結果,-426/-461領域への特異的結合が明かとなった.(3)短鎖(14-21mer)の合成オリゴヌクレオチドを用いた競合ゲルシフト法にて-439/-452領域への結合が示唆された.(4)この部位の塩基配列はCCAAT-enhancer binding protein β(CEBPB),GATA-2およびEts-1の結合部位コンセンサス配列を有し,これらの関与が示唆された.(5)結合部位塩基配列に変異を導入した合成オリゴヌクレオチドを用いて特異性を検定するとともに,抗体によるsupershift法にて,結合蛋白の同定を進めている.\n2. FR遺伝子調節領域特異的結合因子の細胞株特異的発現:(1)前述した特異的結合蛋白がFR発現の程度に関係することを種々の細胞株を用いて検討した.(2)FR高発現細胞株として,KB細胞(親株),Hela細胞,およびメソトレキセート(MTX)耐性KB細胞株を薬剤除去media内で長期培養して得たC2revertant(C2-Rev,FR高発現)を用いた.FR低発現細胞株として,MTX耐性KB細胞株(C2-MTXR),乳癌細胞株(MCF-7),腎細胞株(MDCK),および腎癌細胞株(ACHN,SN12C,およびTK-10)を用いた.(3)FR高発現細胞株として用いたすべての細胞株で特異的結合が認められた.一方,FRの発現が高度に抑制された細胞株C2-MTX-Rでは特異的結合は著しく低下し,さらにFR低発現細胞株すべてにおいてもその結合は低下した.(4)薬剤耐性獲得にともなうFRの発現抑制細胞(C2-MTXR)およびFR低発現細胞株の両方にFR遺伝子発現調節領域への結合蛋白の結合の変化が認められたこと,さらにはC2-Revにおいて可逆的結合の再現が認められたことは,この領域でのFR発現調節が高次的または組織特異的発現調節に関与していることが示唆された.","subitem_description_type":"Abstract"},{"subitem_description":"1. I-dentification of the nuclear DNA-binding proteins involving the specific expression of alpha human folate receptor (hFR) gene : we have identified two promoters which are independently active in a tissue-specific manner in regulation of the alpha hFR gene expression. To further characterize the regulation of these promoters, the transcriptional elements involved in the promoter located upstream of exon 1 were investigated in the transport-defective methotrexate-resistant KB cells (02) by sequence analysis of the promoter, gel shift assays, nuclear run-on assays, and RNase protection assays. Compared to wtKB cells grown in physiologic concentrations of folate, we demonstrate that (1) the 02 cells expressed 2% of a hFR protein and its mRNA ; (2) the transcription rate of a hFR gene was reduced 7 fold in the 02 cells relative to wtKB cells ; (3) the nuclear protein(s) that forms a complex with a -352/-461 (109 bp) DNA fragment located approximately 400 bp upstream of the transcription start site was demonstrated in wtKB cells and was significantly reduced in the C2 cells ; (4) based on competitive gel shift assays using several synthetic oligonucleotides corresponding - 426/-461 DNA sequences, a -4391-452 DNA fragment contained the specific binding sites of this nuclear protein(s). This sequence contains potential binding sites of CCAAT-enhancer binding protein beta, GATA-2, and Ets-1. The supershift assays using antibodies against these proteins will be performed to characterize the factor(s) to this region.\n2. Tissue specific expression of the nuclear factor(s) : the binding of nuclear protein(s) to this DNAfragment was also decreased in MCF-7, MDCK, ACHN, SN12C, and TK- 10 celllines in which expression of the alpha hFR is not detectable. These results suggest that this factor may be involved in the tissue-specific expression of alpha hFR gene and in the modulation of receptor expression in antifolate resistant KB cells.","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:09670793, 研究期間(年度):1997-1998","subitem_description_type":"Other"},{"subitem_description":"出典：研究課題「葉酸受容体の組織特異的遺伝子発現に関与する転写因子の同定とcDNAクローニング」課題番号09670793\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-09670793/096707931998kenkyu_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":"10.24517/00056959","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=60283107"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=60283107","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670793/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670793/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-09670793/096707931998kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-09670793/096707931998kenkyu_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":"2022-05-20"}],"displaytype":"detail","filename":"ME-PR-SAIKAWA-Y-kaken 1999-3p.pdf","filesize":[{"value":"103.5 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-SAIKAWA-Y-kaken 1999-3p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/50650/files/ME-PR-SAIKAWA-Y-kaken 1999-3p.pdf"},"version_id":"5b30d509-9aec-449e-8aa8-f76f2e629d3d"}]},"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":"葉酸受容体の組織特異的遺伝子発現に関与する転写因子の同定とcDNAクローニング","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"葉酸受容体の組織特異的遺伝子発現に関与する転写因子の同定とcDNAクローニング"},{"subitem_title":"Identification and cDNA cloning of the transcriptional factors regulating the tissue-specific expression of human alpha-folate receptor gene","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2835"],"pubdate":{"attribute_name":"公開日","attribute_value":"2022-05-20"},"publish_date":"2022-05-20","publish_status":"0","recid":"50650","relation_version_is_last":true,"title":["葉酸受容体の組織特異的遺伝子発現に関与する転写因子の同定とcDNAクローニング"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T13:04:45.072430+00:00"}