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著作: Goto Tomohiro/[松井 好人]/Fernandes Russell J/Hanson Dennis A/Kubo Ttakahiro/[油形 公則]/Michigami Toshimi/Komori Toshihisa/Fujita Takashi/Yang Liu/Eyre David R/[安井 夏生]/Sp1 Family of Transcription Factors Regulates the Human 2(XI) Collagen Gene (COL11A2) in Saos-2 Osteoblastic Cells./[Journal of Bone and Mineral Research]

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EID
166273
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657228
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2012年9月3日(月) 13:22:19
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大家 隆弘
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松井 好人
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種別 必須 学術論文(審査論文)
言語 必須 英語
招待 推奨
審査 推奨
カテゴリ 推奨
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学究種別 推奨
組織 推奨
  1. 徳島大学.大学院ヘルスバイオサイエンス研究部(2004年4月1日〜2015年3月31日)
著者 必須
  1. (英) Goto Tomohiro
    役割 任意
    貢献度 任意
    学籍番号 推奨
  2. 松井 好人
    役割 任意
    貢献度 任意
    学籍番号 推奨
  3. (英) Fernandes Russell J
    役割 任意
    貢献度 任意
    学籍番号 推奨
  4. (英) Hanson Dennis A
    役割 任意
    貢献度 任意
    学籍番号 推奨
  5. (英) Kubo Ttakahiro
    役割 任意
    貢献度 任意
    学籍番号 推奨
  6. 油形 公則
    役割 任意
    貢献度 任意
    学籍番号 推奨
  7. (英) Michigami Toshimi
    役割 任意
    貢献度 任意
    学籍番号 推奨
  8. (英) Komori Toshihisa
    役割 任意
    貢献度 任意
    学籍番号 推奨
  9. (英) Fujita Takashi
    役割 任意
    貢献度 任意
    学籍番号 推奨
  10. (英) Yang Liu
    役割 任意
    貢献度 任意
    学籍番号 推奨
  11. (英) Eyre David R
    役割 任意
    貢献度 任意
    学籍番号 推奨
  12. 安井 夏生
    役割 任意
    貢献度 任意
    学籍番号 推奨
題名 必須

(英) Sp1 Family of Transcription Factors Regulates the Human 2(XI) Collagen Gene (COL11A2) in Saos-2 Osteoblastic Cells.

副題 任意
要約 任意

(英) Genes encoding type XI collagen, normally associated with chondrogenesis, are also expressed by osteoblasts. By studying Saos-2 cells, we showed that the transcription factors, Sp1, Sp3, and Sp7 (Osterix), regulate COL11A2 expression through its proximal promoter. The findings indicate both ubiquitous and osteoblast-specific mechanisms of collagen gene regulation. Type XI collagen is essential for skeletal morphogenesis. Collagen XI gene regulation has been studied in chondrocytes but not in osteoblasts. We cultured Saos-2 cells, a human osteosarcoma-derived line of osteoblasts, and analyzed them for alpha2(XI) protein and COL11A2 regulatory mechanisms. Although types I and V were the dominant collagens deposited by Saos-2 cells, they expressed COL11A2 mRNA, and alpha2(XI) chains were present in the extracellular matrix. The COL11A2 promoter region (from -149 to -40) containing three Sp1 binding sites was required for promoter activity in transient transfection assays. All three Sp1 sites were critical for binding by nuclear proteins in electrophoretic mobility shift assays. Further analysis using consensus oligonucleotides and specific antibodies as well as chromatin immunoprecipitation assay implicated Sp1 and Sp3 in binding to this promoter region. Overexpressing Sp1 or Sp3 significantly increased COL11A2 promoter activity and endogenous COL11A2 gene expression, an effect that was suppressed by the Sp1-binding inhibitor mithramycin A. Further experiments showed that Sp1, Sp3, CREB-binding protein (CBP), p300, and histone deacetylase (HDAC) were physically associated and HDAC inhibitors (trichostatin A or NaB) upregulated COL11A2 promoter activity and endogenous gene expression. Another Sp1 family member, Sp7 (Osterix), was expressed in Saos-2 cells, but not in chondrocytes, and was shown by chromatin immunoprecipitation to occupy the COL11A2 promoter. Overexpressing Sp7 increased COL11A2 promoter activity and endogenous gene expression, an effect also blocked by mithramycin A. Using siRNA to knockdown Sp1, Sp3, or Sp7, it was shown that depression of any of them decreased COL11A2 promoter activity and endogenous gene expression. Finally, primary cultures of osteoblasts expressed COL11A2 and Sp7, upregulated COL11A2 promoter activity and endogenous gene expression when Sp1, Sp3, or Sp7 were overexpressed, and downregulated them when Sp1, Sp3, or Sp7 were selectively depressed. The results establish that Sp1 proteins regulate COL11A2 transcription by binding to its proximal promoter and directly interacting with CBP, p300, and HDAC.

キーワード 推奨
  1. (英) Base Sequence
  2. (英) Binding Sites
  3. (英) Blotting, Western
  4. (英) Cell Line
  5. (英) Collagen Type XI
  6. (英) DNA Primers
  7. (英) Electrophoretic Mobility Shift Assay
  8. (英) Gene Expression Regulation
  9. (英) Histone Deacetylase Inhibitors
  10. (英) Humans
  11. (英) Immunoprecipitation
  12. (英) Plasmids
  13. (英) Promoter Regions, Genetic
  14. (英) RNA, Messenger
  15. (英) Sp1 Transcription Factor
発行所 推奨
誌名 必須 Journal of Bone and Mineral Research([米国骨代謝学会])
(pISSN: 0884-0431, eISSN: 1523-4681)
ISSN 任意 0884-0431
ISSN: 0884-0431 (pISSN: 0884-0431, eISSN: 1523-4681)
Title: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
Title(ISO): J Bone Miner Res
Supplier: American Society for Bone and Mineral Research
Publisher: Wiley
 (NLM Catalog  (Wiley  (Scopus  (CrossRef (Scopus information is found. [need login])
必須 21
必須 5
必須 661 673
都市 任意
年月日 必須 2006年 5月 初日
URL 任意
DOI 任意 10.1359/jbmr.020605    (→Scopusで検索)
PMID 任意 16734381    (→Scopusで検索)
NAID 任意
WOS 任意
Scopus 任意
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被引用数 任意
指導教員 推奨
備考 任意
  1. (英) Article.Affiliation: Department of Orthopaedics, Institute of Health Biosciences, The University of Tokushima Graduate School, Japan.

  2. (英) Article.PublicationTypeList.PublicationType: Journal Article

  3. (英) Article.PublicationTypeList.PublicationType: Research Support, N.I.H., Extramural