{"_buckets": {"deposit": "6690dc2b-e79f-4df0-935c-5357362ff7e0"}, "_deposit": {"created_by": 1, "id": "1048", "owners": [1], "pid": {"revision_id": 0, "type": "depid", "value": "1048"}, "status": "published"}, "_oai": {"id": "oai:ir.soken.ac.jp:00001048", "sets": ["20"]}, "author_link": ["0", "0", "0"], "item_1_biblio_info_21": {"attribute_name": "書誌情報（ソート用）", "attribute_value_mlt": [{"bibliographicIssueDates": {"bibliographicIssueDate": "2008-03-19", "bibliographicIssueDateType": "Issued"}, "bibliographic_titles": [{}]}]}, "item_1_creator_2": {"attribute_name": "著者名", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "宮本, 貴史"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_1_creator_3": {"attribute_name": "フリガナ", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "ミヤモト, タカシ"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_1_date_granted_11": {"attribute_name": "学位授与年月日", "attribute_value_mlt": [{"subitem_dategranted": "2008-03-19"}]}, "item_1_degree_grantor_5": {"attribute_name": "学位授与機関", "attribute_value_mlt": [{"subitem_degreegrantor": [{"subitem_degreegrantor_name": "総合研究大学院大学"}]}]}, "item_1_degree_name_6": {"attribute_name": "学位名", "attribute_value_mlt": [{"subitem_degreename": "博士（理学）"}]}, "item_1_description_1": {"attribute_name": "ID", "attribute_value_mlt": [{"subitem_description": "2008059", "subitem_description_type": "Other"}]}, "item_1_description_12": {"attribute_name": "要旨", "attribute_value_mlt": [{"subitem_description": "　　It is well known that transcription elongation and repair of UV－damage is coupled.\u003cbr /\u003eIn eukaryotes，stalled RNA pol II recruits several repair proteins to from a complex\u003cbr /\u003econtaining RNA pol II，TFIIH，and the repair proteins．In prokaryotes，stalled RNAP\u003cbr /\u003erecruits Mfd protein．Mfd has a domain homologous to the domain in UvrB which\u003cbr /\u003erecognizes a wide range of structurally diverse lesions in addition to a domain for\u003cbr /\u003einteraction with RNAP．Mfd binds to the DNA segment immediately upstream of the\u003cbr /\u003estalled RNAP and pushes it towards downstream by using the energy of\u003cbr /\u003eATP-hydrolysis to dissociate it from DNA，and speculated to recruit UvrA to perform\u003cbr /\u003ethe repair similar to the global nuclear excision repair. These mechanisms provide the\u003cbr /\u003econcept on the triggering DNA repair rather than that on transcriptional regulation．\u003cbr /\u003e　　Although RNAP tends to stall at all kinds of UV-damages，the efficiency of stallat at \u003cbr /\u003ea UV-damage has not been measured．Therefore，it is possible that inhibition of\u003cbr /\u003etranscription is carried out by a mechanism other than the stall of RNAP．The stall\u003cbr /\u003ecould be overcome by a read-through of the damaged base by misincorporation，\u003cbr /\u003eslippage，Jump，and switching the template strand．These considerations indicate the\u003cbr /\u003emechanism of coulpling of UV-damages and initiation，because initiation generally has\u003cbr /\u003ethe largest dynamic range of transcriptional regulation．Irrespective of the efforts to\u003cbr /\u003efind specific factors triggering the inhibition in initiation，such factors have not been\u003cbr /\u003efound yet, suggesting an alternative possibility that the triggering function has been\u003cbr /\u003ealready installed in RNAP．\u003cbr /\u003e　　Transcription initiation is biochemically separated into several steps．RNAP in the\u003cbr /\u003eform of holoenzyme binds to a promoter and then DNA duplex in the complex is\u003cbr /\u003epartially unwound to expose the template strand of DNA by mostly unknown\u003cbr /\u003emechanism. The resultant complex is called open complex and this complex was once\u003cbr /\u003econsidered to be homogeneous．However，it is now known to involve two or more\u003cbr /\u003econformations which have different catalytic properties，forming branched pathway\u003cbr /\u003emechanism. One branch leads to produce the full-length transcripts（productive\u003cbr /\u003epathway），While another leads to produce only short transcripts iteratively\u003cbr /\u003esynthesized by“moribund complex”which is defined as a complex that produces only\u003cbr /\u003eabortive but no full-length transcripts（non-productive pathway）.On some promoters，\u003cbr /\u003emoribund complex is converted into dead-end complex which still maintains transcript\u003cbr /\u003ebut lacks elongation activity．Accumulation of dead-end complex cause less\u003cbr /\u003estoichiometric synthesis of the full-length products in a single-round transcription\u003cbr /\u003eassay.\u003cbr /\u003e　　To examine the potential coupling and its mechanism，I have studied on the\u003cbr /\u003erelationship between transcription initiation and UV-damage by using purified\u003cbr /\u003ereconstitution system of E.coli．This well-characterized system allows me to consider\u003cbr /\u003ethe relationship between the structure and the function．The system is composed of\u003cbr /\u003eRNAP holoenzyme，a promoter DNA，and four NTPs as substrates．I selected the T7A1\u003cbr /\u003epromoter because little moribund and dead-end complexes are accumulating on this\u003cbr /\u003epromoter，simplifying the analysis of the branched pathway. By irradiating the\u003cbr /\u003epromoter DNA with UV light，UV damages are generated at adjacent pyrimidine bases\u003cbr /\u003ein the promoter region. If the generated damages are up stream from the transcription\u003cbr /\u003estart site，such damages are nothing to do with elongation pause and thus their effect\u003cbr /\u003eon transcription must be limited to the effect on initiation．\u003cbr /\u003e　　In this study, UV-irradiation of the template DNA was found to enhance abortive\u003cbr /\u003einitiation and to induce dead-end complex，namely enhancing the nonproductive\u003cbr /\u003epathway．The positions of UV-damage in the template DNA in moribund and dead-end\u003cbr /\u003ecomplex was compared with those in the binary complex that contains productive\u003cbr /\u003ecomplex．The pyrimidine dimers at several sites in the upstream from the\u003cbr /\u003etranscription start site，but not all，were enriched in the moribund and dead-end\u003cbr /\u003ecomplexes．indicating that these UV-damaged pyrimidine bases induce these\u003cbr /\u003ecomplexes. These positions of the damages are on both template and non-template\u003cbr /\u003estrands. The induction by the damaged pyrimidine residues are confirmed by the\u003cbr /\u003etranscription on DNA fragments which contain single UV-damageds in the upstream\u003cbr /\u003eregion．Furthermore，I found that adjacent pyrimidine bases are conserved atone or\u003cbr /\u003emore positions of those identified on T7A1 promoter among the majority of the E．coli\u003cbr /\u003epromoters，suggesting that these positions would be UV-sensor motif in promoters．\u003cbr /\u003e", "subitem_description_type": "Other"}]}, "item_1_description_7": {"attribute_name": "学位記番号", "attribute_value_mlt": [{"subitem_description": "総研大甲第1166号", "subitem_description_type": "Other"}]}, "item_1_select_14": {"attribute_name": "所蔵", "attribute_value_mlt": [{"subitem_select_item": "有"}]}, "item_1_select_8": {"attribute_name": "研究科", "attribute_value_mlt": [{"subitem_select_item": "生命科学研究科"}]}, "item_1_select_9": {"attribute_name": "専攻", "attribute_value_mlt": [{"subitem_select_item": "18 遺伝学専攻"}]}, "item_1_text_10": {"attribute_name": "学位授与年度", "attribute_value_mlt": [{"subitem_text_value": "2007"}]}, "item_creator": {"attribute_name": "著者", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "MIYAMOTO, Takashi", "creatorNameLang": "en"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_files": {"attribute_name": "ファイル情報", "attribute_type": "file", "attribute_value_mlt": [{"accessrole": "open_date", "date": [{"dateType": "Available", "dateValue": "2016-02-17"}], "displaytype": "simple", "download_preview_message": "", "file_order": 0, "filename": "甲1166_要旨.pdf", "filesize": [{"value": "270.9 kB"}], "format": "application/pdf", "future_date_message": "", "is_thumbnail": false, "licensetype": "license_11", "mimetype": "application/pdf", "size": 270900.0, "url": {"label": "要旨・審査要旨", "url": "https://ir.soken.ac.jp/record/1048/files/甲1166_要旨.pdf"}, "version_id": "dd385290-e2f5-4420-8df4-e7773081ebce"}]}, "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": "thesis", "resourceuri": "http://purl.org/coar/resource_type/c_46ec"}]}, "item_title": "UV-sensor motifs in promoters", "item_titles": {"attribute_name": "タイトル", "attribute_value_mlt": [{"subitem_title": "UV-sensor motifs in promoters"}, {"subitem_title": "UV-sensor motifs in promoters", "subitem_title_language": "en"}]}, "item_type_id": "1", "owner": "1", "path": ["20"], "permalink_uri": "https://ir.soken.ac.jp/records/1048", "pubdate": {"attribute_name": "公開日", "attribute_value": "2010-02-22"}, "publish_date": "2010-02-22", "publish_status": "0", "recid": "1048", "relation": {}, "relation_version_is_last": true, "title": ["UV-sensor motifs in promoters"], "weko_shared_id": -1}