miRNAと転写因子に注目したSCOT遺伝子の肝臓特異的発現抑制機構の解析

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(1)情報処理学会研究報告 IPSJ SIG Technical Report. Vol.2015-BIO-41 No.4 2015/3/20. miRNA ࡜㌿෗ᅉᏊ࡟ὀ┠ࡋࡓ SCOT 㑇ఏᏊࡢ⫢⮚≉␗ⓗⓎ⌧ᢚไᶵᵓࡢゎᯒ ⏣୰⨾᪩ 㐲⸨ಇᚨ ໭ᾏ㐨኱Ꮫ኱Ꮫ㝔 ᝟ሗ⛉Ꮫ◊✲⛉ ⏕࿨ே㛫᝟ሗ⛉Ꮫᑓᨷ ᝟ሗ⏕≀Ꮫ◊✲ᐊ. ⫼ᬒ. 3.. 㑇ఏᏊⓎ⌧᝟ሗࢆඖ࡟ࡋࡓไᚚ㒊఩ࡢྠᐃ. ⚾ࡓࡕࡢ㌟యࡣ㣗࡭≀࠿ࡽ࢚ࢿࣝࢠ࣮ࢆᚓ࡚⏕άࡋ࡚࠾. SCOT 㑇ఏᏊ࡜ఝࡓⓎ⌧ࣃࢱ࣮ࣥࢆ♧ࡍ㑇ఏᏊࢆᢳฟࡍࡿ. ࡾ,୰ᚰⓗ࡞ᙺ๭ࢆᢸ࠺ࡢࡀ⢾࡛࠶ࡿࡀ, ⢾ࡀḞஈࡋࡓ≧. ࡓࡵ, NCBI GEO ࡼࡾࣄࢺ 36 ⤌⧊ࡈ࡜ࡢࡢⓎ⌧ࢹ࣮ࢱࢭࢵ. ែ,ࡘࡲࡾ✵⭡᫬࡟ࡣ, ⬡⫫ࢆཎᩱ࡜ࡋ⫢⮚࡟࠾࠸࡚⏕ᡂ. ࢺ(GSE2361)ࢆྲྀᚓࡋ, Bioconductor Genefilter ࢆ⏝࠸࡚඲. ࡉࢀࡿࢣࢺࣥయࡀ௦࢚᭰ࢿࣝࢠ࣮※࡜ࡋ࡚౑⏝ࡉࢀࡿ. ⫢. ⤌⧊ࡢⓎ⌧ࢆඖ࡟ SCOT 㑇ఏᏊ࡜ఝࡓ 50 㑇ఏᏊࢆᢳฟࡋ. እ⤌⧊࡟࠾࠸࡚ࢧࢡࢩࢽࣝ-CoA:3 ࢣࢺ㓟 CoA ࢺࣛࣥࢫࣇ. ࡓ. ࡑࡢᚋ, ⫢⮚࡟࠾ࡅࡿⓎ⌧್ࡀ௚ࡢ⤌⧊ࡢᖹᆒࡢ 3 ಸ. ࢙࣮ࣛࢮ㸦SCOT㸧㑇ఏᏊ࡟ࡼࡾࢥ࣮ࢻࡉࢀࡿ㓝⣲࡟ࡼࡾ. ௨ୖ࡛࠶ࡿ 12 㑇ఏᏊࢆ⏝࠸, Ward ἲ࡟ࡼࡾࢡࣛࢫࢱࣜࣥ. ศゎࡉࢀࡿࡇ࡜࡛࢚ࢿࣝࢠ࣮࡜࡞ࡿ. ࡑࡢࡓࡵ,SCOT ࡣ⫢. ࢢࢆ⾜ࡗࡓ. ࡲࡓ, ✵⭡᫬࡟࠾ࡅࡿ㑇ఏᏊࡢⓎ⌧ኚືࢆほ. እ⤌⧊࡛ᗈࡃⓎ⌧ࡋ࡚࠸ࡿࡀ, ⫢⮚࡟࠾࠸࡚ࡣ≉␗ⓗ࡟Ⓨ. ᐹࡍࡿࡓࡵ, NCBI GEO ࡼࡾ, ࣐࢘ࢫࡢ✵⭡᫬㛫ࡈ࡜ࡢ⬻,. ⌧ࡀᢚไࡉࢀ࡚࠸ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࡀ, ᮍࡔࡑࡢヲ⣽. ⫢⮚, ⭠, ⭈⮚, ➽⫗࡟࠾ࡅࡿⓎ⌧ࢆㄪ࡭ࡓ GSE24504 ࢆྲྀ. ࡞ᶵᵓࡣ᫂ࡽ࠿࡟ࡉࢀ࡚࠸࡞࠸. ㏆ᖺ࡛ࡣ, ࣐࢘ࢫࢆ⏝࠸. ᚓࡋ, ᭷ពỈ‽ 5%࡛ T ᳨ᐃࢆ⾜ࡗࡓ.. ࡓ ◊ ✲ ࡟ ࡼ ࡾ mir-122 ࡟ ࡼ ࡿ ⩻ ヂ ᢚ ไ , H3K27me3 ࡜ H3K9ac ࡟ࡼࡿࢡ࣐ࣟࢳࣥᵓ㐀ࡢኚ໬[1]ࡢ㛵୚ࡀ♧၀ࡉࢀ ࡚࠸ࡿ. ࡲࡓ, SCOT 㑇ఏᏊࡢⓎ⌧࡟ࡣࣉ࣮ࣟࣔࢱ࣮㡿ᇦ࡟. ⤖ᯝ࡜⪃ᐹ 1. SCOT 㑇ఏᏊࡢᣢࡘᶵ⬟ࡣ့ங㢮࡟࠾࠸࡚ಖᏑ. Ꮡᅾࡍࡿ 2 ࡘࡢ GC-BOX ࡟ࡑࢀࡒࢀ㌿෗ᅉᏊ Sp-1 ࡀ⤖ྜ. ࣐ࣝࢳࣉࣝ࢔ࣛ࢖࣓ࣥࢺࡢ⤖ᯝ, ≉࡟ၥ㢟ࡢ࠶ࡿ㓄ิࡣᏑ. ࡍࡿࡇ࡜ࡀ㔜せ࡛࠶ࡾ, ࡑࡢୖὶࡢ≉ᐃ㡿ᇦࡀⓎ⌧ᢚไ࡟. ᅾࡏࡎ, ⏕≀✀㛫࡟࠾࠸࡚ࡼࡃಖᏑࡉࢀ࡚࠸ࡓ. ࡲࡓ, ࢔. ῝ࡃ㛵ࢃࡗ࡚࠸ࡿ࡜ࡢሗ࿌[2]ࡶᏑᅾࡍࡿ.. ࣑ࣀ㓟㓄ิࢆ⏝࠸ࡓ⣔⤫ᶞࡼࡾ, ࡝ࡢࣀ࣮ࢻ࡟࠾࠸࡚ࡶ. ◊✲ࡢ┠ⓗ SCOT 㑇ఏᏊࡣ⏕యෆ࡟࠾࠸࡚ᚲ㡲࡞ᙺ๭ࢆᣢࡕ, ⫢⮚࡛. Bootstrap ್ࡀ㧗ࡃ, ࡲࡓᯞࡣ㢮ఝࡋ࡚࠸ࡓࡓࡵ, ⏕≀✀࡟ ࡼࡗ࡚≉ᚩⓗ࡞㐍໬ࡣ㉳ࡁ࡚࠾ࡽࡎ, ࡲࡓ, ့ங㢮࡟࠾࠸ ࡚ࡣࡑࡢ㓄ิࡀࡼࡾⰋࡃಖᏑࡉࢀ࡚࠸ࡓ.. ࡢ≉␗ⓗ࡞Ⓨ⌧ᢚไࡀ▱ࡽࢀ࡚࠸ࡿࡀࡑࡢᶵᵓࡣ᫂ࡽ࠿࡟ ࡉࢀ࡚࠸࡞࠸. ࡑࡇ࡛, ᮏ◊✲ࡢ┠ⓗࡣඛ⾜◊✲࡛㛵୚ࡀ ♧၀ࡉࢀ࡚࠸ࡓ miRNA ࡜㌿෗ᅉᏊ࡟╔┠ࢆࡋ, SCOT 㑇ఏ Ꮚࡢ⫢⮚≉␗ⓗ࡞Ⓨ⌧ᶵᵓࡢゎ᫂ࢆࡍࡿࡇ࡜࡜ࡋࡓ.. ᮦᩱ࣭᪉ἲ 1.. SCOT 㑇ఏᏊࡢ့ங㢮㛫࡛ࡢಖᏑᛶࡢホ౯. SCOT ࡢ࢔࣑ࣀ㓟ཬࡧሷᇶ㓄ิࡣ NCBI ࡼࡾ Homo sapiens, Pongo abelii, Macaca mulatta, Mus musculus, Rattus norvegicus, Equus caballus, Sus scrofa, Bos taurus, Gallus gallus, Xenopus laevis ࡢ⏕≀✀࡟ࡘ࠸࡚ྲྀᚓࡋࡓ. ࢔ࣛ࢖ ࣓ࣥࢺ࡟ࡣ clustalX2 ࢆ,⣔⤫ᶞࡢసᡂ࡟ࡣ NJ ἲࢆ౑⏝ࡋ, Bootstrap ್ 1000 ᅇ ࡜ ࡋ ࡓ . ࡲ ࡓ , Window ゎ ᯒ ࡟ ࡣ JCoDA[3]ࢆ౑⏝ࡋࡓ. 2.. ࣄࢺ mir-122 ࡜ SCOT 㑇ఏᏊࡢ⤖ྜ࡟ࡘ࠸࡚ࡢ᳨ウ. mir-122 㓄 ิ ࡣ microrna. org ࡼ ࡾ Homo sapiens, Mus musculus ࡟ࡘ࠸࡚ྲྀᚓࡋࡓ. Alu 㓄ิࡣ Repeatmasker ࡼࡾࣄ ࢺࢤࣀ࣒(GRCh37. 73)୰ࡼࡾྲྀᚓࡋࡓ. miRNA ⤖ྜண ࡟ ࡣ Target Scan, Target Spy, mirwolk, miRDB, MicroCosm ࡢ 5 ✀ࢆ⏝࠸ࡓ. ࢔ࣛ࢖࣓ࣥࢺ࡟ࡣ ClustalX2 ࢆ⏝࠸ࡓ.. ᅗ 1 SCOT ࢔࣑ࣀ㓟㓄ิࢆ⏝࠸ࡓ㐍໬⣔⤫ᶞ NJ ἲ࡛సᡂࡋ, ྛࣀ࣮ࢻ࡟ࡣ 1000 ᅇࢧࣥࣉࣜࣥࢢ࡟ᇶ࡙ࡃ Bootstrap ್, ྛ ᯞ࡟ࡣࡑࡢᯞ㛗ࢆグ㍕ࡋࡓ.. ⣽࠿࡞㐪࠸ࢆぢࡿࡓࡵ࡟ Window ゎᯒࢆ⾜ࡗࡓࡀ, ࡑࡢ㑇 ఏᏊࡢᶵ⬟࡟ཬࡰࡍ㡿ᇦ࡟࠾࠸࡚, 㑇ఏᏊࡢᶵ⬟࡟ᙳ㡪ࢆ ཬࡰࡍࡼ࠺࡞㓄ิࡢኚ໬ࡣ࡞࠿ࡗࡓ. SCOT 㑇ఏᏊࡢ㓄ิ ࡜ᶵ⬟ࡣ့ங㢮㛫࡟࠾࠸࡚ࡼࡃಖᏑࡉࢀ࡚࠸ࡓ. ࡋࡓࡀࡗ ࡚, Ⓨ⌧ไᚚᶵᵓࡶ㢮ఝࡋ࡚࠸ࡿ࡜⪃࠼ࡽࢀ, 㢮ఝࡋࡓⓎ ⌧ࢆ♧ࡍ㑇ఏᏊୖ࡛ඹ㏻ࡋ࡚Ꮡᅾࡍࡿไᚚ㓄ิࡣࡑࡢ㑇ఏ. ⓒ 2015 Information Processing Society of Japan. 1.

(2) 情報処理学会研究報告 IPSJ SIG Technical Report. Vol.2015-BIO-41 No.4 2015/3/20. ᏊࡢⓎ⌧ไᚚ࡟㛵୚ࡋ࡚࠸ࡿྍ⬟ᛶࡀ㧗࠸࡜⪃࠼ࡿࡇ࡜ࡀ. Biomart ࡟ࡼࡾྲྀᚓࡋࡓᚋ, ࡑࢀࡽ㑇ఏᏊୖὶ࡟ඹ㏻ࡋ࡚. ฟ᮶ࡿ.. Ꮡᅾࡍࡿไᚚ㓄ิࢆ TRANSFAC ࡟ࡼࡾ᥈⣴ࡋࡓ. ᮲௳࡜. ࣄࢺ mir-122 ࡣ SCOTmRNA ࡟⤖ྜࡍࡿྍ⬟ᛶࡀప࠸. ࡋ࡚(1)Homo sapiens ࡢ 7 㑇ఏᏊୖὶ࡟Ꮡᅾ(2)Mus musculus. Thorrez et al. 2011 ࡟࠾࠸࡚, Mus musculus ࡢ mir-122 ࡣ. ࡜ Homo sapiens ࡢ SCOT 㑇ఏᏊୖὶ࡟Ꮡᅾ(3)⫢⮚࡛ࡢⓎ⌧. 2.. SCOT ࡢ mRNA 3021-3026 ࡟Ꮡᅾࡍࡿ seed 㓄ิ࡜ࡢ┦⿵ᛶ. ࡢ 3 ࡘ࡜ᐃ⩏ࡋࡓ. ᮲௳ࢆ‶ࡓࡍ㌿෗ᅉᏊ࡜ࡋ࡚ C/EBP. ࡟ᇶ࡙ࡁ⤖ྜࡋ, SCOT ࡢ⩻ヂ㜼ᐖࢆ⾜࠺࡜ࡉࢀ࡚࠸ࡓ. ࡋ. gamma, RXR-alpha, HNF-4alpha, YY1,AP-1 ࢆᚓࡿࡇ࡜ࡀฟ. ࠿ࡋ࡞ࡀࡽ, Homo sapiens ࡟࠾࠸࡚ࡑࡢ㒊఩ࢆ࣌࢔࣡࢖ࢬ. ᮶ࡓ. ≉࡟, HNF-4alpha ࡣ᪂⢾⏕ཬࡧ⬡㉁௦ㅰࡢ࣐ࢫࢱ࣮. ࢔ࣛ࢖࣓ࣥࢺ࡟ࡼࡾẚ㍑ࡋࡓ࡜ࡇࢁ, 㓄ิࡀಖᏑࡉࢀ࡚࠾. 㑇ఏᏊ[4]࡜▱ࡽࢀ, YY1 ࡣ࣑ࢺࢥࣥࢻࣜ࢔࡛ാࡃ㑇ఏᏊࡢ. ࡽࡎ, ࡉࡽ࡟, miRNA ࡢ⤖ྜ㒊఩ண ࡟࠾࠸࡚ࡶ 5 ✀ࡢࢯ. Ⓨ⌧ไᚚࢆᢸ࠺[5]ࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ. ࡲࡓ, ࡇࢀࡽ㌿෗. ࣇ ࢺ ࢘ ࢙ ࢔ ඲ ࡚ ࡛ ⤖ ྜ ࡢ ྍ ⬟ ᛶ ࡀ ྰ ᐃ ࡉ ࢀ ࡓ . Homo. ᅉᏊࡢ✵⭡᫬࡟࠾ࡅࡿⓎ⌧ኚືࢆ᭷ពỈ‽ 5%᳨࡛ᐃࡋࡓ. sapiens ࡟࠾࠸࡚ seed 㓄ิ࡜ࡼࡾ㢮ఝࡋࡓ㓄ิࡀ mRNA. ⤖ᯝ, ⫢⮚࡟࠾࠸࡚ Yy1 ࡀ✵⭡᫬࡟᭷ព࡟Ⓨ⌧ࡀୖ᪼ࡋ࡚. 3436-3441 ࡟Ꮡᅾࡋ࡚࠸ࡓ. 㟋㛗㢮≉␗ⓗ࡟Ꮡᅾࡍࡿ Alu 㓄. ࠸ࡿࡇ࡜ࡀ♧ࡉࢀ, ✵⭡᫬ࡢ㑇ఏᏊไᚚ࡟㛵୚ࡋ࡚࠸ࡿ஦. ิࡣ, miRNA ࡀ⤖ྜࡍࡿ mRNA ࡢ 3'㠀⩻ヂ㡿ᇦ࡟࠾࠸࡚,. ࡀ♧၀ࡉࢀࡓ. ࡲࡓ, Yy1 ࡣ Oct4 ࡜ඹ࡟ࣄࢫࢺ࣓ࣥࢳࣝ໬. ஫࠸࡟ᑐᛂࡏࡎศᕸࡋ࡚࠸ࡿ[4]࡜࠸࠺ࡇ࡜ࡀሗ࿌ࡉࢀ࡚. 㓝⣲ࢆㄏᑟࡍࡿࡇ࡜[6]ࡸ Ctcf ࡜ඹ࡟ Hnf4-alpha ࡸ Polϩࡢ. ࠸ࡿ. SCOT 㑇ఏᏊ࡟࠾ࡅࡿ mRNA ୖࡢศᕸࢆㄪ࡭ࡓࡿ࡜,. Ⓨ⌧࡟ᙳ㡪ࢆ୚࠼ࡿࡇ࡜[7]ࡀ▱ࡽࢀ࡚࠸ࡿ. ௨ୖࡢࡇ࡜ࡼ. ☜࠿࡟ Alu 㢮ఝ㓄ิࡀ 3008-3324 ࡟Ꮡᅾࡋ࡚࠸ࡓ.. ࡾ, SCOT 㑇ఏᏊࡢⓎ⌧ไᚚࣔࢹࣝࢆసᡂࡋࡓ.. ᅗ 2 SCOT mRNA ࡛ࡢ఩⨨㛵ಀ. ᅗ 4 SCOT 㑇ఏᏊࡢⓎ⌧ไᚚࣔࢹࣝ. NCBI ࡼࡾྲྀᚓࡋࡓ NM_000436.3 ࡟ᑐᛂࡋࡓ఩⨨ࢆ♧ࡋ࡚࠶ࡿ.. SCOT ࡟࠾࠸࡚ࡣ SP1 ࡀ⤖ྜࡍࡿࡇ࡜࡛㌿෗ࡀάᛶ໬ࡉࢀࡿ. ᢚไ࡟ാࡃ ሙྜ, ྛ㌿෗ᅉᏊࡀ༠ㄪࡋ࣓ࢳࣝ໬㓝⣲ࡸ⬺࢔ࢭࢳࣝ໬㓝⣲ࢆㄏᑟࡋ, ࣄ. 㐍໬ࡢ㐣⛬࡛㟋㛗㢮≉␗ⓗ࡟Ꮡᅾࡍࡿ Alu 㓄ิࡀᤄධࡀ㉳. ࢫࢺࣥಟ㣭ࢆᘬࡁ㉳ࡇࡍࡇ࡜࡛ࢡ࣐ࣟࢳࣥᵓ㐀ࡢኚ໬ࢆᘬࡁ㉳ࡇࡍ.. ࡁࡓࡇ࡜࡛, Ⓨ⌧ไᚚ࡟㛵୚ࡍࡿ㒊఩ࡀ⨨᥮ࡉࢀࡓྍ⬟ᛶ ࡜ඛ⾜◊✲࡛♧၀ࡉࢀ࡚࠸ࡓ mir-122 ࡟ࡼࡿⓎ⌧ᢚไࡣ, SCOT ࡟┤᥋ാࡃࡢ࡛࡞ࡃ, 㛫᥋ⓗ࡟࡞ࡉࢀ࡚࠸ࡿྍ⬟ᛶ ࡢ஧ࡘࡀ⪃࠼ࡽࢀࡿ.. ⤖ㄽ SCOT 㑇ఏᏊࡢᶵ⬟ࡣ့ங㢮࡟࠾࠸࡚ࡼࡃಖᏑࡉࢀ࡚࠾ࡾ. 㢮ఝⓎ⌧ࢆ♧ࡍ㑇ఏᏊୖ࡟ࡣඹ㏻㌿෗ไᚚ㒊఩ࡀᏑᅾ. ࡑࡢㄪ⠇ᶵᵓࡶྠᵝ࡛࠶ࡿ. ඛ⾜◊✲࡟࠾࠸࡚, SCOT 㑇ఏ. GSE2361 ࢆඖ࡟ SCOT ࡜㢮ఝࡋࡓⓎ⌧ࢆ♧ࡍ㑇ఏᏊࢆᢳฟ. ᏊࡢⓎ⌧ᢚไ࡟ࡣ mir-122 ࡢ㛵୚ࡀሗ࿌ࡉࢀ࡚࠸ࡓࡀ, ࣄ. ࡋ, ࣄ࣮ࢺ࣐ࢵࣉࢆసᡂࡋࡓ.. ࢺ࡟࠾࠸࡚ࡣࡑࡢ⤖ྜ㒊఩ࡀᏑᅾࡋ࡞࠸ࡇ࡜࠿ࡽ㛫᥋ⓗ࡞. 3.. 㛵୚ࡀ♧၀ࡉࢀࡓ. 㑇ఏᏊⓎ⌧᝟ሗ࡟ᇶ࡙ࡁ, Ⓨ⌧ไᚚ࡟ 㛵୚ࡋ࡚࠸ࡿྍ⬟ᛶࡢ㧗࠸㌿෗ᅉᏊࢆྠᐃࡋࡓ.. ཧ⪃ᩥ⊩ 1) Lieven thorrez et al : Genome Research21:95-105(2011) 2) K. E.. Orii et al.: Tohoku J. Exp. Med.,215, 227-236(2008). 3) Steinway et al.: BMC Bioinformatics 11, 284 (2010). 4) Dietrich et al.: Am.. J.Physiol. Gastrointest. Liver Physiol. 293,. G585–90 (2007).. ᅗ 3 SCOT 㢮ఝⓎ⌧ࢆ♧ࡍ㑇ఏᏊࡢࣄ࣮ࢺ࣐ࢵࣉ ᯟ ⥺ 㒊 ୖ ࡼ ࡾ , PREPL, ATP6V1H, FAM131A, ATXN,. 5) Perekatt et al.: Proc. Natl. Acad. Sci. U. S. A. 111, 7695–700 (2014) 6) Zhao et al.: Cancer Res. (2003) 7) Zlatanova, J. & Caiafa, P.: J. Cell Sci. 122, 1275–84 (2009).. OXCT1, GUCY1B3, WDR47 ࡛࠶ࡿ. SCOT 㑇ఏᏊࡢ Gene symbol ࡣ OXCT1 ࡛࠶ࡿ.. ࡇࢀࡽ 6 㑇ఏᏊ࡜ SCOT 㑇ఏᏊࡢୖὶ 3000 ሷᇶ㓄ิࢆ. ⓒ 2015 Information Processing Society of Japan. 2.

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