Genetic regulation of tissue-specific Shh expression by long-range enhancers and chromosome dynamics

• Project/Area Number: 19370003
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Genetics/Genome dynamics
• Research Institution: National Institute of Genetics
• Principal Investigator: SHIROISHI Toshihiko National Institute of Genetics, 系統生物研究センター, 教授 (90171058)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000); Fiscal Year 2008: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000); Fiscal Year 2007: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000)
• Keywords: Shh 遺伝子 / 非翻訳保存配列 / 遺伝子発現制御 / 遠隔エンハンサー / 染色体動態 / Shh遺伝子

Research Abstract

マウス発生胚肢芽では、Shh 遺伝子の転写レベルの変化に応じてShh 遺伝子座から遠く離れた遠隔エンハンサーを含むシス配列とShh 翻訳領域の物理的距離が時期・組織特異的にダイナミックに変化する。本研究では、Shh 遺伝子の転写制御に絡む染色体高次構造変化の分子基盤を明らかにし、その生物学的意義を考察した。本研究成果により、マウス肢芽でのShh 遺伝子発現は、エンハンサーとプロモーターの物理的相互作用に加えて染色体テリトリーからのShh 遺伝子座のルーピングアウトという二段階の染色体高次構造の変化によって制御されることが明らかとなった。また、Shh 遺伝子の遠隔および近位エンハンサーの詳細な発現量の比較解析を行い、前者は短期で比較的低レベルの遺伝子発現を、後者は長期間にわたる高レベルの遺伝子発現を制御している可能性が示唆された。このように、染色体の高次構造のダイナミックな変化により、発生関連遺伝子の発現量について精巧な制御が行われていることが明らかとなった。また、本研究では、肢芽以外において、Shh 遺伝子の口腔から腸管の上皮組織での発現を特異的に制御するエンハンサーを新たに同定した。

Research Products

• [Journal Article] A cluster of three long-range enhancers directs regional Shh expression in the epithelial linings (2009)
  • Author(s): Sagai T, Amano T, Tamura M, Mizushina Y, Sumiyama K and Shiroishi T.
  • Journal Title: Development 136 Pages: 1665-1674
  • Peer Reviewed
• [Journal Article] Chromosomal dynamics at the Shh locus: limb bud-specific differential regulation of competence and active transcription (2009)
  • Author(s): Amano T, Sagai T, Tanabe H, Mizushina Y, Nakazawa H, Shiroishi T
  • Journal Title: Dev Cell. 16 Pages: 47-57
  • NAID: 120006576058
  • Peer Reviewed
• [Journal Article] A cluster of three long-range enhancers directs regional Shh expression in the epithelial linings. (2009)
  • Author(s): Sagai T
  • Journal Title: Development (In press)
  • Peer Reviewed
• [Journal Article] Chromosomal dynamics at the Shh locus : limb bud-specific differential regulation of competence and active transcription. (2009)
  • Author(s): Amano T
  • Journal Title: Developmental Cell 16 Pages: 47-57
  • Peer Reviewed
• [Journal Article] Identification and characterization of new long conserved noncoding sequences in vertebrates (2008)
  • Author(s): Sakuraba Y, Kimura T, Masuya H, Noguchi H, Sezutsu H, Takahasi KR, Toyoda A, Fukumura R, Murata T, Sakaki Y, Yamamura M, Wakana S, Noda T, Shiroishi T , Gondo Y
  • Journal Title: Mamm Genome 19 Pages: 703-712
  • Peer Reviewed
• [Journal Article] Identification and characterization of new long conserved noncoding sequences in vertebrates. (2008)
  • Author(s): Sakuraba Y
  • Journal Title: Mammalian Genome 19 Pages: 703-712
  • Peer Reviewed
• [Journal Article] Members of a novel gene family, Gsdm, are expressed exclusively in the epithelium of the skin and gastrointestinal tract in a highly tissue-specific manner. (2007)
  • Author(s): Tamura M.
  • Journal Title: Genomics 89 Pages: 618-629
  • Peer Reviewed
• [Journal Article] A new Gsdma3 mutation affecting anagen phase of first hair cycle. (2007)
  • Author(s): Tanaka S.
  • Journal Title: Biochem Biophys Res Common. 359 Pages: 902-907
  • Peer Reviewed
• [Journal Article] Mutations in the helix termination motif of mouse type I IRS keratin genes impair the assembly of keratin intermediate filament. (2007)
  • Author(s): Tanaka S.
  • Journal Title: Genomics. 90 Pages: 703-711
  • Peer Reviewed
• [Journal Article] Correlation between Shh expression and DNA methylation status of the limb-specific Shh enhancer region during limb regeneration in amphibians. (2007)
  • Author(s): Yakushiji N.
  • Journal Title: Dev Biol. 312 Pages: 171-182
  • Peer Reviewed
• [Presentation] Sonic hedgehog (Shh)遺伝子の発現制御にかかわる染色体ダイナミクス (2009)
  • Author(s): 城石俊彦
  • Organizer: 第5回生命資源研究・支援センターシンポジウム
  • Place of Presentation: 熊本大学
  • Year and Date: 2009-03-03
• [Presentation] Sonic hedgehog (Shh)遺伝子の発現制御にかかわる染色体ダイナミクス (2009)
  • Author(s): 城石俊彦
  • Organizer: 第5 回生命資源研究・支援センターシンポジウム
  • Place of Presentation: 熊本
• [Presentation] Evolutionary conserved non-coding sequence 1Mb away from the Shh coding sequence acts as limb bud-specific Shh enhancer. (2008)
  • Author(s): Shiroishi T.
  • Organizer: The 16^<th> CDB meeting, "Cis-sequence regulation and its evolution" Kobe
  • Place of Presentation: 理化学研究所神戸
  • Year and Date: 2008-09-29
• [Presentation] Two adjacent long-range-enhancers regulate segmental Shh expression in epithelial linings of endoderm. (2008)
  • Author(s): Sagai T
  • Organizer: 第41回日本発生生物学会
  • Place of Presentation: 徳島県郷土文化会館
  • Year and Date: 2008-05-29
• [Presentation] Gene kissing:A remote enhancer-promoter interaction regulates expression of Sonic hedgehog in mouse limb buds (2008)
  • Author(s): Amano T, Sagai T, Tanabe H, Shiroishi T
  • Organizer: The 3rd Asian Chromosome Colloquium
  • Place of Presentation: Osaka, Japan
• [Presentation] Evolutionary conserved non-coding sequence 1Mb away from the Shh coding sequence acts as limb bud-specific Shhenhancer (2008)
  • Author(s): Shiroishi T
  • Organizer: The16thCDBmeeting,"Cis-sequence regulation and its evolution"
  • Place of Presentation: Kobe
• [Presentation] Long-range エンハンサーによるShh 遺伝子の動的転写制御 (2008)
  • Author(s): 天野孝紀、 嵯峨井知子、田辺秀之、水品洋一、中澤博美、城石俊彦
  • Organizer: 第22回モロシヌス研究会
  • Place of Presentation: 東京
• [Presentation] Hemimelic extra-toes (Hx) mutation gains a new cis-regulatory motif in limb-specific Shh enhancer (2008)
  • Author(s): Amano T, Sagai T, Tanabe H, Shiroishi T
  • Organizer: 第41 回日本発生生物学会(ISDB 共催)
  • Place of Presentation: 徳島
• [Presentation] Twoadjacent long-range-enhancers regulate segmental Shh expression in epithelial linings of endoderm (2008)
  • Author(s): Sagai T, Amano T, Tamura M, Mizushina Y, Yamamoto H, Okagaki A, Sumiyama K, Shiroishi T
  • Organizer: 第41 回日本発生生物学会(ISDB 共催)
  • Place of Presentation: 徳島
• [Presentation] Evolutionary conserved non-coding sequence 1Mb away from the Shh coding region acts as limb bud-specific enhancer in mouse (2008)
  • Author(s): Shiroishi T
  • Organizer: The 1st International Symposium of the Biodiversity Global COE Project, "from Genome to Ecosystem"
  • Place of Presentation: Kyoto
• [Presentation] Shh signaling regulated by a highly conserved remote enhancer is essential for the development of posterior oro-pharyngeal apparatus (2007)
  • Author(s): Sagai T, Amano T, Tamura M, Mizushina Y, Yamamoto H, Sumiyama K, Shiroishi T
  • Organizer: 第40 回日本発生生物学会・第59 回日本細胞生物学会合同大会
  • Place of Presentation: 福岡
• [Presentation] Two Step-wise changes in chromosome architecture regulate Shh expression in the mouse limb bud (2007)
  • Author(s): Amano T and Shiroishi T
  • Organizer: 第40回日本発生生物学会・第59 回日本細胞生物学会合同大会
  • Place of Presentation: 福岡
• [Presentation] Shh signaling regulated by a highly conserved remote enhancer is essential for the development of posterior oro-pharyngeal apparatus (2007)
  • Author(s): Sagai T.
  • Organizer: 日本発生生物学会第40回大会
  • Place of Presentation: 福岡
• [Presentation] Step-wise changes in chromosome architecture regulate Shh expression in the mouse limb bud (2007)
  • Author(s): Amano, T.
  • Organizer: 日本発生生物学会第40回大会
  • Place of Presentation: 福岡