Title
哺乳類Ten-eleven translocation protein (TET) 遺伝子ファミリ
ーの分子進化に関する研究( 本文(Fulltext) )
Author(s)
赤堀, 洋道
Report No.(Doctoral
Degree)
博士(医科学) 連創博甲第39号
Issue Date
2017-03-25
Type
博士論文
Version
ETD
URL
http://hdl.handle.net/20.500.12099/56205
※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。့ங㢮
Ten-eleven translocation protein (TET)
㑇ఏᏊࣇ࣑࣮ࣜࡢศᏊ㐍㛵ࡍࡿ◊✲
Molecular evolution of the Ten-eleven translocation
protein (TET) gene family in mammals
2017 ᖺ 3 ᭶
ᒱ㜧ᏛᏛ㝔㐃ྜ⸆་⒪ሗ◊✲⛉
་⒪ሗᏛᑓᨷ
1
ᮏᏛㄽᩥࡣࠊᏛ⾡◊✲㞧ㄅ㍕ࡉࢀࡓḟࡢཎⴭㄽᩥࢆᇶࡋ࡚సᡂࡋࠊᒱ㜧Ꮫ Ꮫ㝔㐃ྜ⸆་⒪ሗ◊✲⛉ᥦฟࡋࡓࡶࡢ࡛࠶ࡿࠋ
Hiromichi Akahori, Stéphane Guindon, Sumio Yoshizaki and Yoshinori Muto. Molecular Evolution of the TET Gene Family in Mammals.
International Journal of Molecular Sciences; 116 (12), 28472-28485 (2015)
ᮏㄽᩥ࠾ࡅࡿཎⴭㄽᩥࡽࡢᘬ⏝ࡣࠊฟ∧♫ࡽࡢチྍ㸦ࢡ࢚ࣜࢸࣈ࣭ࢥࣔࣥࢬ࣭ࣛ ࢭࣥࢫ㸧ࢆᚓࡓࡶࡢ࡛࠶ࡿࠋ
2
┠
┠ḟ
㸯㸬ᗎㄽ ---3 㸰㸬ᮦᩱ᪉ἲ ---7 㸰㸫㸯 㓄ิࡢྲྀᚓࠊከ㔜࣓ࣛࣥࣥࢺཬࡧ⣔⤫ᶞࡢᵓ⠏ ---7 㸰㸫㸰 ⤌ࡳ࠼ࡢ᳨ฟ ---7 㸰㸫㸱 ศᏊ㐍ゎᯒ ---8 㸰㸫㸲 ࢱࣥࣃࢡ㉁3 ḟඖᵓ㐀ࡢྍど ---9 㸰㸫㸳 ࣑ࣀ㓟ࡢᩡᛶ⨨࠾ࡼࡧศᒱᛶ⨨ࡢᐃ㔞 ---9 㸱㸬⤖ᯝ ---10 㸱㸫㸯 ศᏊ㐍⮬↛㑅ᢥࡘ࠸࡚ࡢᴫㄝ ---10 㸱㸫㸰 ့ங㢮TET ࣇ࣑࣮ࣜ㑇ఏᏊ㓄ิ ---11 㸱㸫㸱 TET ࣇ࣑࣮ࣜ㑇ఏᏊࡢṇࡢ㑅ᢥከᵝ ---14 㸱㸫㸲 ṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࡢྠᐃ ---16 㸱㸫㸳 ⣔⤫ἢࡗࡓࢧࢺ≉␗ⓗ㑅ᢥ㐣⛬ࡢኚ ---18 㸱㸫㸴 3 ḟඖᵓ㐀࣐ࢵࣆࣥࢢ ---23 㸱㸫㸵 ࣑ࣀ㓟ࡢᩡᛶ⨨࠾ࡼࡧศᒱᛶ⨨ ---24 㸲㸬⪃ᐹ ---26 㸳㸬⥲ᣓ ---29 㸴㸬ᩥ⊩ ---30 ㅰ㎡ ---34 Appendix ---353
11. ᗎㄽ
⣽⬊ࡣᵝࠎ࡞ศ≧ែࡸ⎔ቃኚᑐᛂࡋ࡚ࠊ㑇ఏሗࡢⓎ⌧ࢆከᵝኚࡉࡏࠊࡲࡓ ⥔ᣢࡋ࡚࠸ࡿࠋ㏆ᖺࠊࡇ࠺ࡋࡓ㑇ఏᏊⓎ⌧ไᚚࡢࡋࡃࡳࡢ୍ࡘࡋ࡚ࠊ࢚ࣆࢪ࢙ࢿࢸࢡ ࢫࡀࡁࡃὀ┠ࡉࢀ࡚࠸ࡿ [1]ࠋ࢚ࣆࢪ࢙ࢿࢸࢡࢫࡣࠊDNA ࡢሷᇶ㓄ิኚࢆకࢃࡎ 㑇ఏሗⓎ⌧ࡀኚࡍࡿ⌧㇟࡛࠶ࡾࠊࡇࡢኚࡣ⣽⬊ࡽ⣽⬊グ᠈࣭⥅ᢎࡉࢀࡿࠋDNA ሷᇶ㓄ิࡀ⣽⬊ࡢศᚋࡶᏊᏞ⣽⬊ཷࡅ⥅ࡀࢀࠊࡉࡽぶࡽᏊ⥅ᢎࡉࢀࡿࡼ࠺ࠊ ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ኚࡶᏊᏞ⣽⬊ఏ࠼ࡽࢀ࡚࠸ࡃࠋ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ኚࡢ ศᏊⓗᇶ┙ࡣࠊDNA ࡸࣄࢫࢺࣥࢱࣥࣃࢡ㉁ࡢᏛಟ㣭࡛࠶ࡿࠋせ࡞Ꮫಟ㣭ࡋ࡚ࠊDNA ࡘ࠸࡚ࡣࢩࢺࢩࣥሷᇶࡢ࣓ࢳࣝࠊࣄࢫࢺࣥࢱࣥࣃࢡ㉁࡛ࡣࢭࢳࣝࠊ࣓ࢳࣝࠊࣜ ࣥ㓟࡞ࡀ▱ࡽࢀ࡚࠸ࡿ㸦ᅗ1㸧ࠋࡇࢀࡽࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ኚࡀ DNA ሷᇶ㓄ิ ࡢኚࡁࡃ␗࡞ࡿⅬࡣࠊࡑࡢྍ㏫ᛶ࠶ࡿࠋ㏻ᖖࠊᏛಟ㣭ࡉࢀࡓDNA ࡸࣄࢫࢺࣥࢱ ࣥࣃࢡ㉁ࡣ⣽⬊ࡽ⣽⬊⥅ᢎࡉࢀࡿࡀࠊࡑࡢಟ㣭ࡣỌஂኚ࡛ࡣ࡞ࡃࠊᾘཤࡍࡿࡇ ࡶ࡛ࡁࡿࠋࡋࡓࡀࡗ࡚ࠊᏛಟ㣭ࡢຍᾘཤࢆ㏻ࡋ࡚ࠊ㠀ᖖᰂ㌾࡞㑇ఏᏊⓎ⌧ไᚚࡀ ྍ⬟࡞ࡿࠋ ᅗ1 ࢚ࣆࢪ࢙ࢿࢸࢡࢫ DNA ࡢ࣓ࢳࣝࡣࠊ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ኚࡢ୰࡛ࡶྂࡃࡽ▱ࡽࢀ࡚࠾ࡾࠊ⣽⳦ ࡽ့ங㢮⮳ࡿࡲ࡛ᗈ⠊ᅖ࡞◊✲ࡀ⾜ࢃࢀ࡚ࡁࡓࠋ࣓ࢳࣝࡣࢩࢺࢩࣥሷᇶࡢࡳ࡛㉳ࡇࡾࠊ ࣓ࢳࣝᇶࡣࢩࢺࢩࣥࡢ5 ຍࡉࢀࡿࠋ့ங㢮࡛ࡣࠊࢩࢺࢩࣥࡢḟࢢࢽࣥࡀ⥆ࡃ CpG 㓄ิࡀせ࡞࣓ࢳࣝᶆⓗ࡛࠶ࡾࠊࡇࡢ㓄ิࡣ㑇ఏᏊࡢࣉ࣮ࣟࣔࢱ࣮㡿ᇦCpG ࣛࣥ ࢻࡋ࡚ࢡࣛࢫࢱ࣮ࢆᙧᡂࡋ࡚࠸ࡿࠋከࡃࡢሙྜࠊࡇࡢࣉ࣮ࣟࣔࢱ࣮㡿ᇦࡀ࣓ࢳࣝࡉࢀ4 ࡚࠸ࡿࠊࡑࡢ㑇ఏᏊࡢⓎ⌧ࡣᢚไࡉࢀࡿഴྥ࠶ࡾࠊ⬺࣓ࢳࣝࡼࡾ㑇ఏᏊⓎ⌧ࡀά ᛶࡉࢀࡿ [1]ࠋ⌧ᅾࡲ࡛ࠊ㓄അᏊᙧᡂࠊ⬇Ⓨ⏕ཬࡧࢤࣀ࣒ๅࡾ㎸ࡳ࡞ࢆྵࡴከᵝ࡞⣽ ⬊⌧㇟࠾࠸࡚ࠊDNA ࡢ࣓ࢳࣝ⬺࣓ࢳࣝࡀࢫࢵࢳࡋ࡚㔜せ࡞ᙺࢆᯝࡓࡍࡇ ࡀ᫂ࡽࡉࢀ࡚ࡁ࡚࠸ࡿ [2, 3]ࠋࡇ࠺ࡋࡓ DNA ࡢ࣓ࢳࣝ≧ែࡢไᚚࡣࠊ」ᩘ✀ࡢ㓝⣲ ࡼࡿ࣓ࢳࣝп⬺࣓ࢳࣝࡢ┦ኚࡼࡗ࡚ᐇ⌧ࡉࢀ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚ࠊࡇࢀࡽࡢ㓝 ⣲ᛂࡢㄪ⠇ࡣࠊከࡃࡢ⏕≀Ꮫⓗ㐣⛬࡛㔜࡞ពࢆᣢࡘ⪃࠼ࡽࢀ࡚ࡁࡓ [4, 5]ࠋDNA ࡢ࣓ࢳࣝ㛵ࢃࡿ≉␗ⓗ㓝⣲ࡋ࡚ࡣࠊ࣓ࢳࣝᇶࢆࢩࢺࢩࣥሷᇶ㌿⛣ࡍࡿ㓝⣲ࡋ࡚ࠊ DNA ࣓ࢳࣝࢺࣛࣥࢫࣇ࢙࣮ࣛࢮࡀ▱ࡽࢀ࡚࠸ࡿࠋࡇࡢ㓝⣲ࡣࠊDNMT1 ࢆ௦⾲ࡋ࡚ 5 ࡘ ࡢ࣍ࣔࣟࢢࡽᡂࡿ㑇ఏᏊࣇ࣑࣮ࣜࢆᵓᡂࡋ࡚࠸ࡿ [4, 6]ࠋᮏ㑇ఏᏊࣇ࣑࣮ࣜࡢ㓝⣲ࡣ ᶵ⬟ศࡀ㐍ࢇ࡛࠾ࡾࠊ㓄അᏊᙧᡂࡢࢤࣀ࣒ๅࡾ㎸ࡳ࡞ከࡃࡢ㐣⛬࡛ࠊࡑࢀࡒࢀࡀ⊂ ⮬ࡢᶵ⬟ࢆ㐙⾜ࡋ࡚࠸ࡿࠋ୍᪉ࠊDNA ⬺࣓ࢳࣝࡢ㓝⣲ⓗᐇయࡣࠊ᫂࡞㒊ศࡶ࠶ࡿࡀࠊ TET (Ten-eleven translocation protein) 㑇ఏᏊࡢ㛵ࡀ☜ᐇどࡉࢀ࡚࠾ࡾࠊ᪤ከᵝ࡞⣽
⬊⌧㇟ࡢไᚚࡢ㛵ࡀ᫂ࡽࡉࢀࡘࡘ࠶ࡿࠋᮏ◊✲࡛ࡣࠊࡇࡢTET 㑇ఏᏊ╔┠ࡋࡓࠋ
TET 㑇ఏᏊࡀࢥ࣮ࢻࡍࡿࢱࣥࣃࢡ㉁ࡣࠊ2 ౯㕲 2 ࢜࢟ࢯࢢࣝࢱࣝ㓟౫Ꮡᛶࡢࢪ࢜࢟ࢩ ࢤࢼ࣮ࢮ࡛࠶ࡿࠋࡑࡢྡ⛠ ” TET ” ࡣࠊ10 ␒ᰁⰍయ㛗⭎ 22 ࣂࣥࢻ 11 ␒ᰁⰍయ㛗⭎ 23
ࣂࣥࢻࡢ㌿ᗙ(t(10;11)(q22;q23))ࢆᫎࡋ࡚࠸ࡿࠋࡇࡢ㌿ᗙࡣࠊᛴᛶ㦵㧊ᛶⓑ⾑ࡸࣜࣥ
ࣃ⌫ᛶⓑ⾑ࡢᝈ⪅ࡢᰁⰍయ࡛ほᐹࡉࢀ࡚࠾ࡾ [7]ࠊ㌿ᗙࡢ⤖ᯝࡋ࡚ MLL-TET1 ⼥ྜ㑇
ఏᏊࡀ⏕ࡌࡿࠋᚋ⏕ື≀ࡢTET ࢱࣥࣃࢡ㉁ࡣࠊTrypanosoma bruceiࡢJBP1ࠊJBP2a ࢱ
ࣥࣃࢡ㉁ࡢ࢜࢟ࢩࢤࢼ࣮ࢮࢻ࣓ࣥࢆ⏝ࡋࡓ Blast ᳨⣴ࡼࡗ࡚ࠊ᭱ึぢࡘࡅࡽࢀࡓ [8]ࠋ᭦࡞ࡿゎᯒࡼࡾࠊᗈ⠊ᅖࡢ┿᰾⏕≀࠾࠸࡚ TET ࢱࣥࣃࢡ㉁ࢆྵࡴከࡃࡢ᰾㓟ಟ㣭 ࢪ࢜࢟ࢩࢤࢼ࣮ࢮࡢࣇ࣑࣮ࣜࡀྠᐃࡉࢀࡓ [7, 9]ࠋᅗ 2 ♧ࡍࡼ࠺ࠊTET ࢱࣥࣃࢡ㉁ ࡣࠊDNA ୰ࡢ 5-࣓ࢳࣝࢩࢺࢩࣥ㸦5mC㸧ࡽ 5-ࣄࢻࣟ࢟ࢩ࣓ࢳࣝࢩࢺࢩࣥ㸦5hmC㸧ࠊ5-ࣇ࢛࣑ࣝࣝࢩࢺࢩࣥ㸦5fC㸧ࠊ5-࢝ࣝ࣎࢟ࢩࣝࢩࢺࢩࣥ㸦5caC㸧ࡢ୍㐃ࡢ㓟ࢆゐ፹ྍ⬟ ࡞ࡇࡀド᫂ࡉࢀ࡚࠸ࡿ [7, 8, 10, 11]ࠋࡉࡽࠊ5caC ཬࡧ 5fC ࡣࢳ࣑ࣥ DNA ࢢࣜࢥࢩࣛ ࣮ࢮࡼࡾษ㝖ࡉࢀࠊ↓ಟ㣭ࡢࢩࢺࢩࣥ⨨ࡁ࠼ࡽࢀࡿ࠸࠺▱ぢ [10]ࡶ࠶ࡿࠋࡇࢀࡽ ࡢ◊✲ᡂᯝࡣࠊ⬟ືⓗ⬺࣓ࢳࣝࢆᢸ࠺せ㓝⣲ࡀTET ࢱࣥࣃࢡ㉁࡛࠶ࡿࡇࡢࡁ࡞᰿ ᣐ࡞ࡿ [7, 12]ࠋ
5 ᅗ
ᅗ2 TET ࢱࣥࣃࢡ㉁ࡀゐ፹ࡍࡿ 5 ࣓ࢳࣝࢩࢺࢩࣥࡢ㓟
࣓ࢳࣝࢩࢺࢩࣥ5mC ࡢ࣓ࢳࣝᇶ CH3ࡢH ࡽ OH ࡢ㓟ࡼࡾ 5hmC ࡞ࡿࠋࡉࡽ㓟ࡉࢀ H
ࡀ2 ࡘእࢀ 5fC ࡞ࡿࠋ⥆ࡅ࡚ H ࡽ OH 㓟ࡉࢀ 5caC ࡞ࡿࠋ5fC ࡸ 5caC ࡣࠊࢳ࣑ࣥ DNA ࢢࣜ ࢥࢩ࣮ࣛࢮ㸦TDG㸧ሷᇶ㝖ཤಟ㸦BER㸧ࡼࡗ࡚ࢩࢺࢩࣥࡉࢀࡿࠋ
့ங㢮࡛ࡣࠊTET1㸪TET2㸪TET3 ࠸࠺ 3 ࡘࡢ TET ࣃࣛࣟࢢࡀྠᐃࡉࢀ࡚࠸ࡿ [7, 9, 12]ࠋ
ᅗ 3 ♧ࡍࡼ࠺ࠊࡑࢀࡒࢀ࢝ࣝ࣎࢟ࢩࣝᮎ➃ゐ፹ࢥࢻ࣓ࣥࢆ᭷ࡋࠊࡇࡢࢥࢻ࣓
ࣥࡣCys-rich domain ࠊࡁ࡞ Low-complexity insert ࡛ศ᩿ࡉࢀࡓ Double-stranded
șhelix (DSBH) domain ࡛ᵓᡂࡉࢀࡿ [7, 13]ࠋLow-complexity insert ࡢ㛗ࡉࡣࠊࡇࢀ
ࡽTET ࣃࣛࣟࢢࡢ㛫࡛ࡁࡃ␗࡞ࡾࠊࡑࡢ㓄ิࡣࢇಖᏑࡉࢀ࡚࠸࡞࠸ [9]ࠋࡲࡓࠊ
TET1 ཬࡧ TET3 ࡣࠊDNA ⤖ྜࣔࢳ࣮ࣇࡋ࡚▱ࡽࢀ࡚࠸ࡿ࣑ࣀᮎ➃ CXXC domain
㸦ᅗ3㸧ࡀᏑᅾࡍࡿ [7, 12]ࠋ୍᪉ࠊTET2 ࡣᰁⰍయ㏫ࡼࡾ TET2 ♽ඛ㑇ఏᏊࡀศࡉ
ࢀ࡚ࠊCXXC ࢻ࣓ࣥࢆࢥ࣮ࢻࡍࡿ㡿ᇦゐ፹ࢻ࣓ࣥࢆࢥ࣮ࢻࡍࡿ㡿ᇦࡀูࠎࡢ㑇ఏ Ꮚ࡞ࡗࡓ [6, 9, 14]ࠋࡉࡽࠊࡇࢀࡽ 3 ࡘࡢ TET 㑇ఏᏊࡣࠊ␗࡞ࡗࡓ⤌⧊≉␗ⓗⓎ⌧ࣃ ࢱ࣮ࣥࢆ♧ࡍ [15, 16]ࠋࡇࡢࡼ࠺ࠊTET 㑇ఏᏊࡣࠊ㑇ఏᏊ㔜」㐺ᛂ㐍ࡀཎᅉ࡛ᶵ ⬟ⓗከᵝࡋࡓࡶࡢ᥎ ࡉࢀࡿࡀࠊࡑࡢ㐍㐣⛬ࡢヲ⣽ࡣࡃゎᯒࡉࢀ࡚࠸࡞࠸ࠋ
6 ᅗ ᅗ3 TET 㑇ఏᏊࡢྛ㡿ᇦ㸦ࢻ࣓ࣥ㸧 ᮏ◊✲࡛ࡣTET 㑇ఏᏊࣇ࣑࣮ࣜࡢศᏊ㐍Ꮫⓗ≉ᛶࢆ᫂ࡽࡍࡿࡓࡵࠊ㑇ఏᏊࣞ ࡛࣋ࣝࡢṇࡢ㑅ᢥࠊࡑࡢ㑅ᢥᅽࡉࡽࡉࢀࡿ࣑ࣀ㓟ࢧࢺࡘ࠸࡚ヲ⣽ゎᯒࡋࡓࠋ ࡇࡢゎᯒࡢ୰࡛ࠊ㑅ᢥࣃࢱ࣮ࣥࡀ㐍⣔⤫࡛ࡣ㐪ࢃ࡞࠸ࡀࠊ࣑ࣀ㓟㓄ิἢࡗ࡚ኚືࡍ ࡿ௬ᐃࡋࡓࢥࢻࣥࣔࢹࣝ㸦ࢧࢺࣔࢹࣝ㸧ࢆ⏝ࡋࡓ [17-20]ࠋࡲࡓࠊ㐍⣔⤫࣑ࣀ 㓟㓄ิࡶኚືࡍࡿ㑅ᢥࣃࢱ࣮ࣥࢆチࡍࢥࢻࣥࣔࢹࣝ㸦ࣈࣛࣥࢳࢧࢺࣔࢹࣝ㸧ࡶ⪃៖ ࡋࡓ [21]ࠋࡇࢀࡽࡢࣔࢹࣝࡼࡿゎᯒࡽࠊ့ங㢮ࡢ TET 㑇ఏᏊࣇ࣑࣮ࣜࡀ TET1 TET2 ࡢ㢖⦾࡞㐺ᛂᨺᩓࢆక࠺ṇࡢ㑅ᢥࢆཷࡅࡓࡇࢆ᫂ࡽࡋࡓࠋࡲࡓࠊTET3 㑇ఏᏊ ࡣ㐍ⓗⴭࡋࡃಖᏑࡉࢀ࡚࠸ࡿࡇࡽࠊࡑࡢ⏕≀Ꮫⓗព⩏ࡘ࠸࡚ࡶ⪃ᐹࡋࡓࠋ
7
22. ᮦᩱ᪉ἲ
2 – 1 㓄ิࡢྲྀᚓࠊከ㔜࣓ࣛࣥࣥࢺཬࡧ⣔⤫ᶞࡢᵓ⠏
TET 㑇ఏᏊࡢ┦ྠ㓄ิࡣࠊMuto et al. [22]ࡢ᪉ἲᚑ࠸ࠊ௨ୗ♧ࡍᡭ㡰࡛㞟ࡋࡓࠋ
᭱ึࠊ့ங㢮ࡢ TET ࣃࣛࣟࢢ࠾ࡼࡧ࣮࢜ࢯࣟࢢࢆ⥙⨶ⓗ㞟ࡍࡿࡓࡵࠊGenBank
(http://www.ncbi.nlm.nih.gov/genbank/)ࢱࣥࣃࢡࢹ࣮ࢱ࣮࣋ࢫࢆ⏝࠸ࠊࣄࢺࡢ TET1ࠊ TET2ࠊTET3 ࡢ࣑ࣀ㓟㓄ิࢆ᳨⣴ࢩ࣮ࢻࡋ࡚ࠊBlastp (http://blast.ncbi.nlm.nih.gov/ Blast.cgi)᳨⣴ࢆ⾜ࡗࡓ [23]ࠋຍ࠼࡚ࠊEnsembl Compara ࢹ࣮ࢱ࣮࣋ࢫ (http://www. ensembl.org/info/genome/compara/index.html) [24]ࢆ⏝࠸ࠊTET1ࠊTET2ࠊTET3ࠊࡢ࣡ ࣮ࢻࢆ᳨⣴ㄒࡋࠊEnsembl API v80 (http://www.ensembl.org/info/docs/api/ index.html)
ࢆ⏝ࡋ࡚TET1ࠊTET2ࠊTET3 ࡢ࣮࢜ࢯࣟࢢࢆ㞟ࡋࡓࠋEnsembl API ࡢࣥࢱ࣮ࣇ
࢙࣮ࢫࡣࠊAppendix 5 ♧ࡍ⊂⮬ࣉࣟࢢ࣒ࣛࢆసᡂࡋ⮬ືࡋࡓࠋྲྀᚓࡋࡓ㓄ิࡽࠊ㔜
」ࡋࡓ㓄ิࡸ࡞㓄ิࢆ㝖እࡋࠊ᭱⤊ⓗ့ங㢮TET 㑇ఏᏊࡢ 130 㓄ิࢆᚓࡓࠋᅇ
ࡢゎᯒ⏝࠸ࡓ130 㓄ิࡘ࠸࡚ࠊ✀ྡࢡࢭࢵࢩࣙࣥࢼࣥࣂ࣮ࢆ Appendix 1 ♧ࡋࡓࠋ TET1ࠊTET2ࠊTET3 ࡑࢀࡒࢀࡢ࣑ࣀ㓟㓄ิࡢከ㔜࣓ࣛࣥࣥࢺ㸦ከ㔜ᩚิ㸧ࡣࠊ iterative refinement method㸦FFT-NS-i㸧࡛᭱㐺ࡉࢀࡓࣃ࣓࣮ࣛࢱタᐃᇶ࡙࠸࡚ࠊ MAFFT ࣉࣟࢢ࣒ࣛ(http://mafft.cbrc.jp/alignment/software/)࡛ᐇ⾜ࡋࡓ [25]ࠋᚓࡽࢀࡓከ 㔜࣓ࣛࣥࣥࢺ㓄ิࡘ࠸࡚ࠊᩚิࡀ༑ศ࡞㓄ิ㡿ᇦࡣࠊtrimAL 1.2 ࣉࣟࢢ࣒ࣛ [26] ࢆ⏝࠸࡚๐㝖ࡋࡓࠋAppendix 2㹼4 ࡣࠊ᭱⤊ⓗ࡞࣑ࣀ㓟㓄ิࡢከ㔜࣓ࣛࣥࣥࢺࢆ ♧ࡍࠋሷᇶ㓄ิࡢከ㔜࣓ࣛࣥࣥࢺࡣࠊ࣑ࣀ㓟ࡢከ㔜࣓ࣛࣥࣥࢺ㓄ิᑐᛂࡍࡿ DNA ሷᇶ㓄ิࡽࠊࢥࢻࣥ༢࡛ PAL2NAL [27]ࣉࣟࢢ࣒ࣛࢆ⏝࠸࡚⏕ᡂࡋࡓࠋ ⣔ ⤫ ᶞ ࡢ ᵓ ⠏ ⏝ ࠸ ࡿ ሷ ᇶ ⨨ ࣔ ࢹ ࣝ ࡣ ࠊjModeltest2 ࣉ ࣟ ࢢ ࣛ ࣒ [28](https:// github.com/ddarriba/jmodeltest2)ࡼࡾࠊAkaike information ᇶ‽ᇶ࡙࠸࡚᥎ᐃࡋࡓࠋ ᭱ᑬἲࡼࡿ⣔⤫ᶞ᥎ᐃࡣࠊPhyML v3.0(https://code.google.com/p/phyml/)ࣉࣟࢢ࣒ࣛ [29]ࢆ⏝ࡋࡓࠋPhyML ࡢࣃ࣓࣮ࣛࢱタᐃࡣࠊapproximate likelihood ratio (aLRT) method ᑐᛂࡋࠊሷᇶ⨨ࣔࢹࣝࡋ࡚ jModeltest2 ࡛᭱㐺᥎ᐃࡉࢀࡓ GTR+I+G4 ࢆ ⏝࠸ࡓࠋ
2 – 2 ⤌ࡳ࠼ࡢ᳨ฟ
TET 㑇ఏᏊࡢከ㔜࣓ࣛࣥࣥࢺ㓄ิෆࡢ㓄ิ㛫ࡢ⤌ࡳ࠼ࡢ᭷↓ࢆㄪࡓࠋࡇࢀࡣ ௨๓ࡢ◊✲ [30]࡛᥇⏝ࡋࡓࠊHYPHY ࣃࢵࢣ࣮ࢪࡢ GARD ࣝࢦࣜࢬ࣒ [31]ࢆ⏝ࡋࡓࠋ
8 ⤌ࡳ࠼࣋ࣥࢺࢆ᳨ฟࡍࡿࡓࡵ࣓ࣛࣥࣥࢺ㓄ิ࡛ࡢሷᇶ᩿∦ࡢ⣔⤫࠾ࡅࡿ୍ ⮴ࢆㄪࡿᡭἲࢆ⏝࠸࡚࠸ࡿࠋࡇࡇ࡛ࡣࠊ㏻ᖖࡢྍ㏫ࣔࢹࣝ㸦GTR㸧ࢆ⏝࠸ࠊ4 ࣮ࣞࢺࢡ ࣛࢫࢆᣢࡘᬑ㐢ⓗ࡞㞳ᩓⓗศᕸࢆ⏝ࡍࡿGARD ゎᯒࢆᐇ⾜ࡋࡓࠋHYPHY ࣃࢵࢣ࣮ࢪࡢ GARDProcess.bf ࣂࢵࢳࣇࣝࡢ୰࡛ࡣࠊ⤫ィⓗ᭷ព࡞⤖ྜࣈࣞࢡ࣏ࣥࢺࢆྠ ᐃࡍࡿࡓࡵKishino-Hasegawa ࢸࢫࢺ [32]ࡀᐇ⾜ࡉࢀࡿࠋ 22 – 3 ศᏊ㐍ゎᯒ
ṇࡢ⮬↛㑅ᢥࡢ᳨ฟࡸ㐍㏿ᗘࡢ᥎ᐃࡣࠊPAML ࣃࢵࢣ࣮ࢪ ver. 4.8 [33]ࡢ codeml ࣉ
ࣟࢢ࣒ࣛ Fitmodel ver. 0.5.3 [21]ࡼࡿ᭱ᑬἲࢆ⏝࠸ࡓࠋcodeml ゎᯒࡣ TET1ࠊ
TET2ࠊTET3 ࡢከ㔜࣓ࣛࣥࣥࢺሷᇶ㓄ิᑐࡋู࡚ࠎᐇࡋࠊྛ㑇ఏᏊࡢࢥࢻࣥࢧ ࢺࡀ␗࡞ࡿȰẚ࡞ࡿࡇࡀྍ⬟࡞ࢧࢺࣔࢹࣝࢆ⏝ࡋࡓ [19, 20]ࠋ᭱ึࡢࣔࢹࣝ࣌ ࡛ࡣࠊᖐ↓௬ㄝࣔࢹࣝM1a ࡣࠊ1 ᮍ‶ࡢȰ0㸯➼ࡋ࠸Ȱ1ࡢ㸰ࡘࡢࢥࢻࣥࢧࢺ࢝ࢸࢦ ࣮ࣜࢆᣢࡗ࡚࠾ࡾࠊࡑࢀࡒࢀ㢖ᗘp0࠾ࡼࡧp1ࡀᑐᛂࡍࡿࠋࡇࡇ࡛ࠊȰࡣdN/dS ࡛࠶ࡾࠊ dN ࡣࠊ㠀ྠ⩏⨨㏿ᗘࢆ⾲ࡋࠊdS ࡣྠ⩏⨨㏿ᗘࢆ⾲ࡍࠋM2a ࡣᑐ❧௬ㄝࣔࢹ࡛ࣝ࠶ࡾࠊ ṇࡢ㑅ᢥࢆ⾲ࡍ1 ௨ୖࡢȰ2ࡑࡢ㢖ᗘp2࠸࠺ࠊ㏣ຍࡢ࢝ࢸࢦ࣮ࣜࢆᣢࡗ࡚࠸ࡿࠋ2 ࡘ┠ ࡢࣔࢹࣝ࣌ࡣࠊM7 M8 ࡛࠶ࡾࠊM7 ࡣࠊȰ್ࡀ 0 1 ࡢ㛫࡛șศᕸᚑ࠺ᖐ↓௬ㄝ ࣔࢹࣝࢆ⾲ࡋ࡚࠸ࡿࠋࡇࡇ࡛șࣃ࣓࣮ࣛࢱࡣࠊș(p,q)࡛࠶ࡿࠋ୍᪉ࠊM8 ࡣࠊᑐ❧௬ㄝࣔࢹ ࡛ࣝ࠶ࡾࠊṇࡢ㑅ᢥࢆチᐜࡍࡿ1 ௨ୖࡢȰ1ࡑࡢ㢖ᗘp1࠸࠺ࠊࢥࢻࣥࢧࢺᑐࡍࡿ㏣ ຍࡢ࢝ࢸࢦ࣮ࣜࢆᣢࡗ࡚࠸ࡿࠋྛࣔࢹࣝ࣌ࡢ⤫ィⓗ᭷ពᛶࡢ᳨ドࡣࠊcodeml ࡢฟຊ⤖ ᯝࢆ⏝࠸࡚ᑬᗘẚ᳨ᐃ㸦LRT㸧ࢆ⾜ࡗࡓࠋᑬᗘẚ᳨ᐃࡢ⤫ィ㔞ࡣࠊ2 ࡘࡢࣔࢹࣝ㛫ࡢᑐᩘᑬ ᗘࡢᕪࡢ2 ಸ㸦2ǼlnL = 2(lnL1-lnL0)㸧┦ᙜࡍࡿࠋᖐ↓௬ㄝࡀṇࡋ࠸ࡁࠊLRT ್ࡣ⮬ ⏤ᗘ2 ࡢ࢝ 2 ศᕸ࡞ࡿࠋLRT ್ࡀ㈇ࡢሙྜࡣࠊ␗࡞ࡗࡓึᮇ್Ȱ㸦0.05ࠊ0.1ࠊ0.15㸧 ࢆ⏝࠸࡚codeml ࢆ」ᩘᅇᐇ⾜ࡋࠊᚓࡽࢀࡓᑐᩘᑬᗘࡢ್᭱ࢆ LRT ࡢィ⟬⏝࠸ࡓࠋ⮬ ⏤ᗘᑐᛂࡍࡿ࢝2 ศᕸࡢ್ࡣࠊPAML ࣃࢵࢣ࣮ࢪࡢ࢝ 2 ࣉࣟࢢ࣒ࣛࡽᚓࡓࠋ ṇࡢ㑅ᢥࢆチᐜࡍࡿᑐ❧௬ㄝࣔࢹࣝ㸦M2aࠊM8㸧࠾࠸࡚ࡣࠊ≉ᐃࡢࢥࢻࣥࡀṇࡢ㑅ᢥࢆ ཷࡅ࡚㐍ࡋࡓ࠸࠺ᚋ☜⋡㸦PP㸧ࢆỴᐃࡍࡿࡓࡵࠊBayes Empirical Bayes (BEB) ᡭ ἲࢆ⏝࠸ࡓ [34]ࠋࡑࡢ㝿ࠊᚋ☜⋡ࡢ 0.95 ࢆ࢝ࢵࢺ࢜ࣇ್ࡋ࡚ࠊࡑࢀࡼࡾࡁ࠸ሙྜ ṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺࡋ࡚ྠᐃࡋࡓࠋ
Fitmodel ࣉࣟࢢ࣒ࣛࡣࠊࢥࢻࣥ㐍ࡘ࠸࡚࣐ࣝࢥࣇኚㄪ࣐ࣝࢥࣇࣔࢹࣝ࠶ࡿ࠸ࡣࢫ ࢵࢳࣥࢢࣔࢹࣝࢆ௬ᐃࡋ࡚࠾ࡾ [21]ࠊcodeml ࡣ␗࡞ࡗࡓሗࡀᚓࡽࢀࡿࠋࡑࡇ࡛ࠊTET1ࠊ TET2ࠊTET3 ࣓ࣛࣥࣥࢺ㓄ิࡢࡑࢀࡒࢀࡘ࠸࡚ࠊFitmodel ࣉࣟࢢ࣒࡛ࣛゎᯒࡋࡓࠋ
9 ࢫࢵࢳࣥࢢࣔࢹࣝࡣࠊࢥࢻࣥࢧࢺ㛫࡛ࡢȰኚຍ࠼࡚⣔⤫ἢࡗࡓȰࡢኚືࢆチᐜ ࡍࡿࣔࢹ࡛ࣝ࠶ࡿࠋࡇࡢゎᯒ࡛ࡣࠊM0ࠊM3ࠊ M3 + S1ࠊM3 + S2 ࡢ 4 ࡘࡢࣔࢹࣝࢆ⏝ ࡋࡓࠋM0 ࣔࢹࣝࡣࠊࢥࢻࣥࢧࢺ⣔⤫ࡘ࠸࡚༢୍ࡢȰࢆ௬ᐃࡋࠊM3 ࣔࢹࣝࡣࠊࢥࢻ ࣥࢧࢺࡀ㸱ࡘࡢ␗࡞ࡗࡓȰࡢ್㸦Ȱ1㸺Ȱ2㸺Ȱ3㸧ࢆᣢࡘ௬ᐃࡍࡿࣔࢹ࡛ࣝ࠶ࡿࠋࡉࡽ ࠊ⣔⤫ἢࡗࡓȰࡢኚື㸦ࢫࢵࢳࣥࢢ㸧ࢆ௬ᐃࡋࠊȰ1ࡽȰ2ࠊȰ1ࡽȰ3ࠊȰ2ࡽ Ȱ3ࡢࡼ࠺࡞Ȱ㛫ࡢࢫࢵࢳࣥࢢ࣮ࣞࢺࡀ➼ࡋ࠸ᐃࡉࢀࡿࡣࠊࡑࡢࢫࢵࢳࣥࢢࣔࢹ ࣝࢆM3 + S1 ࣔࢹࣝࡧࠊ➼ࡋࡃ࡞࠸ࡁࡣ M3 + S2 ࣔࢹࣝࡪࠋࡑࡋ࡚ࠊM0 M3ࠊM3 M3 + S1ࠊM3 + S1 M3 + S2 ࡢࠊ2 ࡘࡢࣔࢹࣝࡢࡑࢀࡒࢀࡢ㛫ࡘ࠸࡚ᑬᗘ ẚ᳨ᐃࢆᐇ⾜ࡋࡓࠋᑐᩘᑬᗘࡢ㐪࠸ࡢ᭷ពᛶࢆỴᐃࡍࡿࡓࡵࡣࠊ࢝2 ศᕸࢆ⏝࠸ࡓࠋ ᮏ◊✲࡛ࡣࠊFitmodel ࣉࣟࢢ࣒ࣛࡢᐇ⾜⤖ᯝࡢ࠺ࡕࠊྛࢥࢻࣥ࠾ࡼࡧࣈࣛࣥࢳ࡛ࡢ 3 ␒┠ ࡢȰࢡࣛࢫ㸦Ȱ3㸧ࡢᚋ☜⋡ࢆホ౯ࡋࠊࡑࡢฟຊ⤖ᯝࢆAppendix 6 ♧ࡍ⊂⮬సᡂࡋ ࡓperl ࣉࣟࢢ࣒ࣛࢆ⏝࠸࡚ฎ⌮ࡋࡓࠋ ௨ୖࡢࠊDatamonkey ࢙࢘ࣈࢧ࣮ࣂ࣮ [35]࡛ᥦ౪ࡉࢀࡿࠊ5 ࡘࡢ᭱ᑬἲࣉࣟࢢ࣒ࣛ ࢆ⏝࠸࡚ࠊࢧࢺࣔࢹࣝࡼࡿṇࡢ㑅ᢥࢆゎᯒࡋࡓࠋ⏝ࡋࡓࣉࣟࢢ࣒ࣛࡣࠊFixed Effect Likelihood㸦FEL㸧ࠊFast Unbiased Bayesian AppRoximation㸦FUBAR㸧 [18]ࠊMixed Effects Model of Evolution㸦MEME㸧 [36]ࠊSingle Likelihood Ancestor Counting㸦SLAC㸧ࠊ Random Effect Likelihood㸦REL㸧 [17]࡛࠶ࡿࠋṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻ࡛ࣥ࠶ࡿྠᐃࡍ ࡿᇶ‽ࢆࠊFUBAR ἲ࡛ࡣᚋ☜⋡ࡀ 0.9 ࡼࡾࡶࡁ࠸ࠊFELࠊMEMEࠊSLAC ἲ࡛ࡣ࢝
ࢵࢺ࢜ࣇP ್ࡀ 0.1 ௨ୗࠊࡲࡓ REL ἲ࡛ࡣ࣋ࢬࣇࢡࢱ࣮ࡀ 50 ࡼࡾࡁ࠸ࡶࡢࡋࡓࠋ
22 – 4 ࢱࣥࣃࢡ㉁ ḟඖᵓ㐀ࡢྍど
TET ゐ፹ࢻ࣓ࣥࡢ 3 ḟඖᵓ㐀ࡢྍどࠊࡑࡢ 3 ḟඖᵓ㐀ୖࡢṇࡢ㑅ᢥࢆཷࡅࡓࢧ
ࢺࡢ࣐ࢵࣆࣥࢢࡣࠊPyMOL ศᏊࢢࣛࣇࢵࢡࢫࢩࢫࢸ࣒ ver.1.3 ࢆ⏝ࡋࡓࠋࡲࡓࠊTET
ࢱࣥࣃࢡ㉁ࡢࣄࢺ࡛ࡢኚ␗ሗࡣࠊUniProt ࢹ࣮ࢱ࣮࣋ࢫ [37]ࡼࡾධᡭࡋࡓࠋ 2 – 5 ࣑ࣀ㓟ࡢᩡᛶ⨨࠾ࡼࡧศᒱᛶ⨨ࡢᐃ㔞 TET1ࠊTET2ࠊTET3 ࡢࡑࢀࡒࢀࡢ㑇ఏᏊࡘ࠸࡚ࠊᩡᛶࡢ࣑ࣀ㓟⨨ཬࡧศᒱᛶ ࡢ࣑ࣀ㓟⨨ࢆ᥎ ࡋࡓࠋࡇࢀࡣࠊcodeMLancestral ࣉࣟࢢ࣒ࣛࣃࢵࢣ࣮ࢪࢆ⏝࠸ࡿ Castoe ࡽ [38]ࡢ᪉ἲࢆ⏝ࡋࡓࠋcodeMLancestral ࣉࣟࢢ࣒࡛ࣛࡣࠊJTT ࣑ࣀ㓟⨨ࣔ ࢹࣝࡢୗ࡛ࠊᩡᛶ࠾ࡼࡧศᒱᛶࡢ࣑ࣀ㓟⨨ࡘ࠸࡚⣔⤫ᶞࡢࣈࣛࣥࢳ࣌ࡘ࠸ ࡚ ྛ ࣑ ࣀ 㓟 ࢧ ࢺ ࡢ ᚋ ☜ ⋡ ࡢ ್ ࢆ ᥎ ᐃ ࡋ ࡓ ࠋ ྛ ⣔ ⤫ ᶞ ࡢ ࣈ ࣛ ࣥ ࢳ ࡢ 㛗 ࡉ ࡣ
10 PhyML(ver3.0)ࢆ⏝ࡋ࡚᥎ ࡋࡓࠋࡲࡓࠊAppendix 7 ♧ࡍ⊂⮬ࡢ Perl ࣉࣟࢢ࣒ࣛࢆ⏝ ࠸࡚ࠊಶࠎࡢ࣑ࣀ㓟ࢧࢺࡢᩡᛶ࠾ࡼࡧศᒱᛶ⨨ࡢ☜⋡ࢆ࡚ࡢࣈࣛࣥࢳ࣌ࡘ ࠸࡚ྜ⟬ࡋࡓࠋࡇࡢྜ⟬್ࢆ⏝࠸࡚ࠊྛ࣑ࣀ㓟ࢧࢺࡢᩡᛶ⨨ࡢ☜⋡ศᒱᛶ⨨ ࡢ☜⋡ࡢ㛵ಀࢆࣉࣟࢵࢺࡋࡓᅗࢆసᡂࡋࡓࠋ
33. ⤖ᯝ
3 – 1 ศᏊ㐍⮬↛㑅ᢥࡘ࠸࡚ࡢᴫㄝ ᭱ึࠊᮏ◊✲ࡢせࢸ࣮࣐࡛࠶ࡿศᏊ㐍ࡸ⮬↛㑅ᢥࡢᴫᛕࡘ࠸࡚⡆༢グ㏙ࡍࡿࠋ ୍⯡ࠊศᏊ㐍ࡣDNA ሷᇶ㓄ิࡢኚࡸࢱࣥࣃࢡ㉁ࡢ࣑ࣀ㓟㓄ิࡢኚࡋ࡚ᤊ࠼ࡿ ࡇࡀ࡛ࡁࡿࡀࠊ⏕≀ࡢ㐍ࡣࡇࡢศᏊ㐍ᇶ࡙࠸࡚࠸ࡿࠋࡋࡓࡀࡗ࡚ࠊศᏊ㐍ࡘ ࠸࡚ࡢ▱㆑ࡣࠊ⏕≀㐍ࡸ⏕≀࠶ࡿ࠸ࡣ⏕యศᏊࡑࡢࡶࡢࡢ⌮ゎࡁࡃ㈉⊩࡛ࡁࡿࠋ ศᏊ㐍ゎᯒࡽᚓࡽࢀࡿሗࡢ୰࡛ࠊ᭱ࡶ㔜せ࡞ᴫᛕࡢ୍ࡘࡀࠕ⮬↛㑅ᢥ࡛ࠖ࠶ࡿࠋ ⮬↛㑅ᢥࡣࠊࢳ࣮ࣕࣝࢬ ࢲ࣮࢘ࣥࡀ⏕≀㐍ࡢ࣓࢝ࢽࢬ࣒ࡋ࡚ึࡵ࡚ᥦࡋ࡚௨᮶ࠊ ⌧ᅾࡲ࡛ࡑࡢ㔜せᛶࡀ☜❧ࡉࢀ࡚ࡁࡓࠋᥦᙜึࠊ⮬↛㑅ᢥࡢᴫᛕࡣ⏕≀ಶయࡢࡳࢆᑐ ㇟ࡋ࡚࠸ࡓࡀࠊ⌧ᅾ࡛ࡣ㑇ఏᏊࡸ࣑ࣀ㓟࡛ࣞ࣋ࣝࡢ⮬↛㑅ᢥࡶ㔜せどࡉࢀ࡚࠸ࡿࠋ㏻ ᖖࠊ⮬↛㑅ᢥࡣࠕṇࡢ㑅ᢥࠖࠕ㈇ࡢ㑅ᢥࠖ࠸࠺ࡘࡢࢱࣉࡀ༊ูࡉࢀࡿࠋṇࡢ㑅 ᢥࡣࠊ⏕Ꮡ᭷࡞ኚ␗㸦ኚ㸧ࡀ㞟ᅋ୰ᛴ㏿ᗈࡀࡗ࡚࠸ࡃ᪉ྥാࡃ⮬↛㑅ᢥ࡛ࠊ 㐺ᛂ㐍ࡢࡋࡃࡳࡢᇶ♏࡞ࡿࠋ㑇ఏᏊࡸࢱࣥࣃࢡ㉁ࡢ࡛ࣞ࣋ࣝࡣࠊṇࡢ㑅ᢥࡀస⏝ࡍࡿ ࡑࡢ㓄ิࡀᛴ㏿ኚ㸦㐍㸧ࡋ࡚࠸ࡃࠋ୍᪉ࠊ㈇ࡢ㑅ᢥࡣࠊ᭷ᐖ࡞ኚ␗ࡀ㞟ᅋࡽ㝖 ཤࡉࢀࡿ᪉ྥാࡃ⮬↛㑅ᢥ࡛࠶ࡿࠋࡇࡢሙྜࠊࢱࣥࣃࢡ㉁ࡢ࣑ࣀ㓟㓄ิࡣ࠶ࡲࡾኚ ࡏࡎࠊⴭࡋࡃಖᏑࡉࢀࡿഴྥ࠶ࡿࠋᮏ◊✲࡛ࡣ≉ṇࡢ㑅ᢥࡢ᳨ฟ╔┠ࡋ࡚࠸ࡿࡀࠊ ṇࡢ㑅ᢥࢆཷࡅࡓ㑇ఏᏊࡢ࣑ࣀ㓟ࢧࢺࡣࠊࡑࡢ㑇ఏᏊࡢ≉Ṧࡸᶵ⬟ศ㔜せ࡞ᙺ ࢆᯝࡓࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋ ᮏ◊✲࡛⏝࠸ࡓcodeml ࡸ Fitmodel ࡞ࡢࣉࣟࢢ࣒ࣛࡣࠊȰࡢ᥎ᐃࡼࡗ࡚ṇࡢ㑅ᢥࢆ ᳨ฟࡋ࡚࠸ࡿࠋ୍⯡ࠊDNA ࢆᵓᡂࡍࡿ AࠊGࠊCࠊT ࡢ 4 ࡘࡢሷᇶࡢ࠺ࡕࠊ୪ࢇࡔ 3 ࡘ ࡢሷᇶࡀࢥࢻࣥ࡞ࡾ1 ࡘࡢ࣑ࣀ㓟ࢆࢥ࣮ࢻࡍࡿࡀࠊ43=64 ㏻ࡾࡢ୪ࡧ᪉ࡀ࠶ࡿࡢ࡛ࠊ1 ࡘࡢ࣑ࣀ㓟」ᩘࡢࢥࢻࣥࡀᑐᛂࡍࡿሙྜࡀከ࠸ࠋࡋࡓࡀࡗ࡚ࠊ⏕≀㐍ࡢ㐣⛬࠾࠸ ࡚ࠊሷᇶࡀ⨨ࡉࢀࡓࡁࢥ࣮ࢻࡍࡿ࣑ࣀ㓟ࡀྠࡌሙྜኚࡍࡿሙྜࡀ⏕ࡌࠊ๓⪅ࢆ ྠ⩏⨨ࠊᚋ⪅ࢆ㠀ྠ⩏⨨࠸࠺ࠋྠ⩏⨨ࡢሙྜࡣࠊ࣑ࣀ㓟ࡀྠࡌ࡞ࡢ࡛ྠࡌࢱࣥ ࣃࢡ㉁ࡀ⏕ᡂࡉࢀࡿࡀࠊ㠀ྠ⩏⨨ࡢሙྜࡣࠊ࣑ࣀ㓟ࡀ␗࡞ࡿࡢ࡛␗࡞ࡿࢱࣥࣃࢡ㉁ࡀ ⏕ᡂࡉࢀࡿࠋ๓㏙ࡢࡼ࠺ࠊȰࡣ㠀ྠ⩏⨨㏿ᗘࢆྠ⩏⨨㏿ᗘ࡛㝖ࡋࡓ್࡛࠶ࡿ㸦Ȱ =11 dN/dS㸧ࠋࡋࡓࡀࡗ࡚ࠊȰ>1 ࡢሙྜࡣࠊሷᇶࡢ⨨ࡼࡾ࣑ࣀ㓟ࡀᛴ㏿ኚࡋ࡚࠸ࡿࡇ ࢆពࡋ࡚࠾ࡾࠊṇࡢ㑅ᢥࢆཷࡅࡓࡇࡀ᫂ࡽ࡞ࡿࠋࡲࡓࠊȰ<1 ࡢሙྜࡣࠊሷᇶࡢ ⨨ࡣ࠶ࡗࡓࡋ࡚ࡶࠊ࣑ࣀ㓟ࡣኚࡋ࡞࠸≧ែ࡛ಖᣢࡉࢀ࡚ࡁࡓࡇࡽࠊ㈇ࡢ㑅ᢥ ࡀാ࠸࡚࠸ࡿࡇࡀ♧ࡉࢀࡿࠋȰࡢ᥎ᐃࡼࡿ⮬↛㑅ᢥࡢ᳨ฟἲពእࡶከࡃࡢᡭἲࡀ▱ ࡽࢀ࡚࠸ࡿࡀࠊȰࡢ᥎ᐃࡣ㑇ఏᏊࡢሷᇶ㓄ิࡢࡳࡽ᥎ᐃ࡛ࡁࡿ࠸࠺౽ᛶࡽࠊከࡃ ࡢ◊✲࡛ࢃࢀ࡚࠸ࡿࠋ 33 – 2 ့ங㢮 7(7 ࣇ࣑࣮ࣜ㑇ఏᏊ㓄ิ
GenBank ࢹ࣮ࢱ࣮࣋ࢫ࠾ࡅࡿࣄࢺ TET ࣑ࣀ㓟㓄ิࢆ᳨⣴ࢩ࣮ࢻࡋࡓ Blastp ᳨⣴
ࠊEnsemble Compara ࢹ࣮ࢱ࣮࣋ࢫ࡛ࡢ Ensambl API ࣉࣟࢢ࣒ࣛࡢ⏝ࡼࡾࠊ့ங
㢮࡛130 ࡢ TET 㑇ఏᏊ┦ྠ㓄ิࢆ☜ㄆ࡛ࡁࡓࠋAppendix 1 ♧ࡋࡓࡼ࠺ࠊࡑࢀࡒࢀࡢ 㑇ఏᏊ࡛ࠊ့ங㢮ࡢᑡ࡞ࡃࡶ39 ⏕≀✀ࡢ┦ྠ㓄ิࢆධᡭฟ᮶ࡓࠋࡇࡢࡼ࠺࡞ᗈ⠊࡞✀࡛ TET1ࠊTET2ࠊTET3 ࡢ┦ྠ㓄ิࡀぢ࠸ࡔࡉࢀࡿࡇࡣࠊ့ங㢮ࡢከᵝ௨๓ TET 㑇ఏ Ꮚࡀ2 ᅇࡢ㐃⥆ࡍࡿ㑇ఏᏊ㔜」ࢆ⤒㦂ࡋ࡚࠸ࡿࡇࢆᙉࡃ♧၀ࡍࡿ [7, 9]ࠋࡇࢀࡽ TET 㑇 ఏᏊࡢࣃࣛࣟࢢ㛫࡛ࡣࠊ㓄ิࡢ㢮ఝᛶࡀゐ፹㡿ᇦ௨እࡣⴭࡋࡃప࠸ࡓࡵࠊࣃࣛࣟࢢࡢከ 㔜࣓ࣛࣥࣥࢺࡣྍ⬟࡛࠶ࡗࡓࠋࡑࡇ࡛ࠊᅇࡢ◊✲࡛ࡣࡑࢀࡒࢀࡢࣃࣛࣟࢢࢆูࠎ ࣓ࣛࣥࣥࢺࡋ࡚ゎᯒࡋࡓࠋ Anisimova ࡽࡣ [39]ࠊ⤌ࡳ࠼ࡀṇࡢ㑅ᢥࡢ᳨ฟࡁࡃᙳ㡪ࡍࡿࡇࢆ♧ࡋࡓࠋࡑࡇ ࡛ࠊ᭱ึ⤌ࡳ࠼ࣈࣞࢡ࣏ࣥࢺࡢ᭷↓ࢆ3 ࡘࡢ TET ࣃࣛࣟࢢࡢ࣓ࣛࣥࣥࢺ㓄ิ ࡘ࠸࡚ゎᯒࡋࡓࠋゎᯒࡣࠊHyPhy ࣃࢵࢣ࣮ࢪ࡛ᐇ⾜ࡉࢀࡿ㑇ఏⓗࣝࢦࣜࢬ࣒ࡼࡿ ⤌ࡳ࠼᳨ฟ㸦GARD㸧ࢶ࣮ࣝࢆ⏝ࡋࡓ [31]ࠋTET1ࠊTET2ࠊTET3 ࡢྛ㑇ఏᏊ࠾࠸ ࡚ࠊp ್ࡀ 0.01 ࡛ࡢ Kishino-Hasegawa㸦KH㸧ࢸࢫࢺ࡛ࡣ⤌ࡳ࠼ࡣ᳨ฟࡉࢀ࡞ࡗࡓࠋ ḟࠊTET1ࠊTET2ࠊTET3 㑇ఏᏊࡢሷᇶ㓄ิከ㔜࣓ࣛࣥࣥࢺࢆ⏝࠸࡚ࠊPhyML ࣉ ࣟࢢ࣒࡛ࣛ᭱ᑬἲࡼࡿ⣔⤫ᶞࢆᵓ⠏ࡋࡓࠋࡇࢀࡽࡢ⣔⤫ᶞࢆࠊᘬࡁ⥆ࡃ㐍ゎᯒ⏝࠸ ࡓࠋᅗ4ࠊᅗ 5ࠊᅗ 6 ࡣࠊTET1ࠊTET2ࠊTET3 㑇ఏᏊࡢࡑࢀࡒࢀࡢ⣔⤫ᶞࢆ♧ࡋࡓࠋ
12 ᅗ
ᅗ4 TET1 ⣔⤫ᶞ
ሷᇶ㓄ิከ㔜࣓ࣛࣥࣥࢺࢆ⏝࠸࡚ࠊPhyML ࣉࣟࢢ࣒࡛ࣛᵓ⠏ࡋࡓ᭱ᑬ⣔⤫ᶞࠋFitmodel ゎᯒ⤖ᯝࡢ ࠺ࡕࠊPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࢥࢻࣥࢧࢺᩘࡀ90 ࣃ࣮ࢭࣥࢱࣝ௨ୖࡢࣈࣛࣥࢳࢆ㟷Ⰽ࡛♧ࡍࠋ
13 ᅗ
ᅗ5 TET2 ⣔⤫ᶞ
ሷᇶ㓄ิከ㔜࣓ࣛࣥࣥࢺࢆ⏝࠸࡚ࠊPhyML ࣉࣟࢢ࣒࡛ࣛᵓ⠏ࡋࡓ᭱ᑬ⣔⤫ᶞࠋFitmodel ゎᯒ⤖ᯝࡢ ࠺ࡕࠊPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࢥࢻࣥࢧࢺᩘࡀ90 ࣃ࣮ࢭࣥࢱࣝ௨ୖࡢࣈࣛࣥࢳࢆ࢜ࣞࣥࢪⰍ࡛♧ࡍࠋ
14 ᅗ ᅗ6 TET3 ⣔⤫ᶞ ሷᇶ㓄ิከ㔜࣓ࣛࣥࣥࢺࢆ⏝࠸࡚ࠊPhyML ࣉࣟࢢ࣒࡛ࣛᵓ⠏ࡋࡓ᭱ᑬ⣔⤫ᶞࠋFitmodel ゎᯒ⤖ᯝࡢ ࠺ࡕࠊPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࢥࢻࣥࢧࢺᩘࡀ90 ࣃ࣮ࢭࣥࢱࣝ௨ୖࡢࣈࣛࣥࢳࢆ⥳Ⰽ࡛♧ࡍࠋ 3 – 3 TET ࣇ࣑࣮ࣜ㑇ఏᏊࡢṇࡢ㑅ᢥከᵝ ⾲1 ࡣࠊcodeml ࣉࣟࢢ࣒ࣛ [19, 20]ࡢฟຊ⤖ᯝࢆせ⣙ࡋࡓࡶࡢ࡛࠶ࡿࠋ᭱ึࡢ M0 ࣔࢹ ࣝࡣࠊྛ࣓ࣛࣥࣥࢺ㓄ิ࠸࡚ࠊࡍ࡚ࡢࢥࢻࣥࢧࢺ୍ᚊࡢȰࡢ್ࢆᐃࡍࡿ ࣔࢹ࡛ࣝ࠶ࡿࠋࡋࡓࡀࡗ࡚ࠊM0 ࣔࢹࣝࡢ⤖ᯝࡣࠊTET 㑇ఏᏊࡢྛࣃࣛࣟࢢ࡛ࡢ⯡ⓗ࡞
15 Ȱࡢ್ࢆ᥎ᐃࡍࡿࠋࡇࡢ⤖ᯝࡼࢀࡤࠊTET1 TET2 ࡛ࡑࢀࡒࢀȰࡀ 0.385ࠊ0.346 㢮 ఝࡋࡓ್࡞ࡾࠊTET3 ࡛ࡣࡇࢀࡽࡼࡾⴭࡋࡃᑠࡉ࠸ 0.115 ࡛࠶ࡗࡓࠋࡋࡋࠊ࠸ࡎࢀࡏ ࡼTET 㑇ఏᏊ࡛ࡣȰࡢ್ࡀ 1 ࡼࡾᑠࡉࡃࠊࢇࡢ⣔⤫࠾ࡼࡧࢧࢺ࠸࡚ࠊ㈇ࡢ㑅 ᢥࢆཷࡅ࡚㐍ࡋࡓࡇࡀ᥎ᐃࡉࢀࡓࠋࡲࡓࠊM2a ཬࡧ M8 ࡣࠊᖐ↓௬ㄝࣔࢹ࡛ࣝ࠶ࡿ M1a ཬࡧ M7 ᑐࡍࡿᑐ❧௬ㄝࣔࢹ࡛ࣝ࠶ࡿࠋࡇࢀࡽࡢࣔࢹࣝࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓ᥎ ᐃࡉࢀࡿ 1 ࡼࡾࡶࡁ࠸Ȱ್ࡢ࢝ࢸࢦࣜࢆᣢࡗ࡚࠾ࡾࠊࡇࡢࣔࢹࣝࡀᡂ❧ࡍࢀࡤࠊࡑࡢ㑇 ఏᏊࡣṇࡢ㑅ᢥࢆཷࡅ࡚࠸ࡿࡇࡀ♧ࡉࢀࡿࠋ⾲୰ࠊM2a ࡛ࡣȰ2ࡀࠊM8 ࡛ࡣȰ1ࡀṇࡢ 㑅ᢥࢆチᐜࡍࡿ࢝ࢸࢦ࡛ࣜ࠶ࡿࠋᅇࡢ⤖ᯝ࡛ࡣࠊTET1 ࡢ M8 ࣔࢹ࡛ࣝȰ1 = 1.295ࠊTET2 ࡛ࡣȰ1 = 1.558 ࡢ್ࢆᚓࡓࠋࡲࡓࠊࡇࡢࣔࢹࣝࡢ p1ࡢ್ࡽࠊTET1 ࡛ࡣ 3.8%ࠊTET2 ࡛ ࡣ 2.2%ࡢࢥࢻࣥࢧࢺࡀ 1 ࡼࡾࡶࡁ࡞್࡞ࡾࠊṇࡢ㑅ᢥࢆཷࡅࡓࡇࡀ᥎ᐃࡉࢀࡓࠋ BEB ᡭἲࡼࡗ࡚ṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࢆ᥎ᐃࡋࡓ⤖ᯝࡣࠊ⾲ࡢ᭱ྑิグ㍕ࡋ ࡓࠋM2a ࣔࢹ࡛ࣝࡣࠊTET 㑇ఏᏊࡢఱࢀࡶṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡿࢧࢺࡣ↓ ࡗࡓࡀࠊM8 ࣔࢹ࡛ࣝࡣࠊTET1 ཬࡧ TET2 ṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡿࢧࢺࡀぢ࠸ ࡔࡉࢀࡓࠋTET1 ࡛ࡣࠊ524Fࠊ811M ࡢ 2 ࣑ࣀ㓟ࠊTET2 ࡛ࡣ 123Rࠊ469Vࠊ673Vࠊ968Pࠊ 1002Tࠊ1205Sࠊ767S ࡢ 7 ࣑ࣀ㓟ࡀヱᙜࡋࠊᚋ☜⋡ࡶ 0.95 ࡼࡾࡁ࠸ࠋTET3 ࡘ࠸ ࡚ࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡿ࣑ࣀ㓟ࢧࢺࡣ↓ࡃࠊ㈇ࡢ㑅ᢥࢆཷࡅ࡚㐍ࡋࡓ ᥎ᐃࡉࢀࡿࢧࢺࡢྜ㸦M1a ࣔࢹࣝࡢ p0㸧ࡀࠊࡇࡢ㑇ఏᏊࡘ࠸࡚ࡣⴭࡋࡃࡁ࠸ࡇ ࡀ᫂ࡽ࡞ࡗࡓ㸦p0 = 0.86㸧ࠋ
16 ⾲
⾲1 PAML ࣉࣟࢢ࣒ࣛࡼࡿ TET 㑇ఏᏊࡢ᭱ᑬἲゎᯒ
Model Parameter estimates a LnL 2ΔlnL (P value) Positively selected sites b,c
TET1: M0 ω = 0.385 -87143.158462 None M1a ω0 = 0.205, p0 = 0.623, ω1 = 1.000, p1 = 0.377 -86007.766228 Not allowed M2a ω0 = 0.205, p0 = 0.623, ω1 = 1.000, p1 = 0.298, ω2 = 1.000, p2 =0.078 -86007.766228 M2a vs. M1a 0 (P=1) None M7 p = 0.679, q = 0.983 -85718.609725 Not allowed M8 p0 = 0.962, p = 0.744, q = 1.197, p1 = 0.038, ω1 = 1.294 -85707.99818 M8 vs. M7 21.22309 (P < 0.001) 524F, 811M TET2: M0 ω = 0.346 -75677.716183 None M1a ω0 = 0.184, p0 = 0.664, ω1 = 1.000, p1 = 0.336 -74593.700860 Not allowed M2a ω0 = 0.184, p0 = 0.664, ω1 = 1.000, p1 = 0.292, ω2 = 1.000, p2 =0.044 -74593.700860 M2a vs. M1a 0 (P=1) None M7 p =0.617, q = 1.064 -74286.497185 Not allowed M8 p0 = 0.978, p = 0.671, q = 1.250, p1 = 0.022, ω1 = 1.558 -74265.863135 M8 vs. M7 41.2681 (P < 0.001) 123R*, 469V, 673V, 968P, 1002T, 1205S, 1767S TET3: M0 ω = 0.115 -48719.653509 None M1a ω0 = 0.056, p0 = 0.860, ω1 = 1.000, p1 = 0.140 -47597.069946 Not allowed M2a ω0 = 0.056, p0 = 0.859, ω1 = 1.000, p1 = 0.139, ω2 = 999.000, p2 =0.002 -47599.818203 M2a vs. M1a -5.496514 (P=1) None M7 p = 0.237, q = 1.380 -47268.934744 Not allowed M8 p0 = 0.999, p = 0.237, q = 1.380, p1 = 0.001, ω1 =999.000 -47270.552239 M8 vs. M7 -3.23499 (P=1) None a : p, q ࡣࠊM7ࠊM8 ࣔࢹࣝ࠾࠸࡚Ȱ್ࢆỴᐃࡍࡿșศᕸࡢࣃ࣓࣮ࣛࢱࠋ b : ᚋ☜⋡ࡀ 0.95 ࡼࡾ ࡁ࠸࣑ࣀ㓟ࢧࢺࢆ♧ࡋࡓࠋ * : ᚋ☜⋡ࡀ 0.99 ࡼࡾࡁ࠸࣑ࣀ㓟ࢧࢺ c :࣑ࣀ㓟⨨ࡢᇶ‽ ࡋ࡚ࠊࡑࢀࡒࢀࣄࢺࡢTET1ࠊTET2ࠊTET3 ࢆ⏝࠸ࡓࠋ 3 – 4 ṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࡢྠᐃ ᮏ◊✲࡛ࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࢆࡼࡾ㧗ឤᗘ᳨ฟࡍࡿࡓࡵࠊPAML ຍ࠼࡚」ᩘࡢࢧࢺࣔࢹࣝࣉࣟࢢ࣒ࣛࢆ⏝࠸࡚ࠊṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺࢆ᳨ฟࡍࡿゎᯒ ࢆ⾜ࡗࡓࠋ⏝ࡋࡓࣉࣟࢢ࣒ࣛࡣࠊPAML ࡢ codeml ௨እ Datamonkey ࢙࢘ࣈࢧ࣮ࣂ࣮ [35]࡛ࡢ FELࠊFUBARࠊMEMEࠊRELࠊSLAC ࡛࠶ࡾࠊ᭱⤊ⓗ 6 ✀ࡢᡭἲࡼࡿ⤖ᯝ ࢆᚓࡓࠋࡇࢀࡽࡢゎᯒ⤖ᯝࡽࠊࡢ◊✲ [40, 41]࠾ࡅࡿุᐃᇶ‽ࢆ⪃៖ࡋ࡚ࠊᑡ࡞ࡃ
17
ࡶ 3 ࡘࡢᡭἲ࡛ṇࡢ㑅ᢥࢆཷࡅࡓྠᐃࡉࢀࡓࢥࢻࣥࢆṇࡢ㑅ᢥࢆཷࡅࡓࡶࡢࡋࡓࠋ
TET1 ࡛ࡣ 34ࠊTET2 ࡛ࡣ 29ࠊTET3 ࡛ࡣ 13 ࡢࢥࢻࣥࡀṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡓࠋ
ྛTET 㑇ఏᏊࡘ࠸࡚ࡢ⤖ᯝࢆࢻ࣓ࣥඹᅗ㸵ࠊᅗ 8ࠊᅗ 9 ♧ࡍࠋTET1 ࡛ṇࡢ㑅 ᢥࡀ᭱ࡶከࡃ᳨ฟࡉࢀࠊࡑࡢከࡃࡣ࣑ࣀᮎ➃ഃࡢࢧࢺ㞟୰ࡋ࡚࠸ࡿ㸦ᅗ㸵㸧ࠋTET2 ࡘ࠸࡚ࡶྠᵝࡢഴྥࡀㄆࡵࡽࢀࠊ᳨ฟࡉࢀࡓࢥࢻࣥࢧࢺࡢᩘࡶྠ⛬ᗘ࡛࠶ࡿ㸦ᅗ㸶㸧ࠋ ୍᪉ࠊTET3 ࡘ࠸࡚ࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺᩘࡣᑡ࡞ࡃࠊ㛗ࢃࡓࡗ࡚ࡰᆒ➼ ศ ᕸ ࡋ ࡚ ࠸ ࡿ 㸦 ᅗ 9 㸧ࠋ ඛ ㏙ ࡢ ࡼ ࠺ ࠊ TET ࣇ ࣑ ࣜ ࣮ ࢱ ࣥ ࣃ ࢡ ㉁ ࡣ ࠊ DSBH ࠊ Low-complexity-InsertࠊCys-rich Insert ࡽᵓᡂࡉࢀࡿ࢝ࣝ࣎࢟ࢩࣝᮎ➃ゐ፹㡿ᇦࢆᣢࡘ ࡀ [13]ࠊ⯆῝࠸ࡇࠊṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࡣ DSBH ཬࡧ Cys-rich Insert ࡣ ࢇᏑᅾࡋ࡞ࡗࡓࠋࡲࡓࠊCXXC domain ࡣ DNA ࡢ CpG 㓄ิ⤖ྜࡍࡿࡀ [7]ࠊࡇࡢ 㡿ᇦࡶṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࡀ୍ࡘࡶ᥎ᐃࡉࢀ࡞ࡗࡓࠋࡇࢀࡣࠊᙉ࠸㈇ࡢ㑅ᢥࡀാ ࠸࡚࠸ࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋࡇࢀᑐࡋ࡚ࠊゐ፹㡿ᇦ࡛ṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡓ ࢥࢻࣥࡣࠊࡑࡢࢇࡀLow-complexity-Insert Ꮡᅾࡋ࡚࠾ࡾࠊ␗࡞ࡗࡓ㑅ᢥᅽࡀࡇࢀ ࡽࡢ㡿ᇦ࡛ാ࠸࡚࠸ࡿࡇࡀ♧ࡉࢀࡓࠋ ᅗ ᅗ7 TET1 㑇ఏᏊࡢṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺ TET1 㑇ఏᏊ࠾ࡅࡿṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࢆ⾲♧ࠋྛࢻ࣓ࣥࡣࠊࣄࢺ TET1 㑇ఏᏊࡢ CXXC domainࠊCys-rich InsertࠊDSBHࠊLow-complexity insert ࢆඖ⾲♧ࡋ࡚࠸ࡿࠋྛ㡿ᇦࡢቃ⏺ࡣࠊ ࣑ࣀ㓟ࡢ␒ྕࡀグࡋ࡚࠶ࡿࠋࡲࡓࠊṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺࢆڸ༳࡛࣑ࣀ㓟␒ྕඹ♧ࡋ࡚࠸ࡿࠋ PAML ゎᯒࡢ M8 ࣔࢹ᳨࡛ࣝฟࡉࢀࡓࢧࢺࡣ㟷Ⰽ࡛♧ࡍࠋ
18 ᅗ
ᅗ8 TET2 㑇ఏᏊࡢṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺ
TET2 㑇ఏᏊ࠾ࡅࡿṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࢆ⾲♧ࠋྛࢻ࣓ࣥࡣࠊࣄࢺ TET2 㑇ఏᏊࡢ Cys-rich InsertࠊDSBHࠊLow-complexity insert ࢆඖ⾲♧ࡋ࡚࠸ࡿࠋྛ㡿ᇦࡢቃ⏺ࡣࠊ࣑ࣀ㓟ࡢ␒ྕࡀグࡋ ࡚࠶ࡿࠋࡲࡓࠊṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺࢆڸ༳࡛࣑ࣀ㓟␒ྕඹ♧ࡋ࡚࠸ࡿࠋPAML ゎᯒࡢ M8 ࣔࢹ ᳨࡛ࣝฟࡉࢀࡓࢧࢺࡣ㟷Ⰽ࡛♧ࡍࠋ
ᅗ9 TET3 㑇ఏᏊࡢṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺ
TET3 㑇ఏᏊ࠾ࡅࡿṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺࢆ⾲♧ࠋྛࢻ࣓ࣥࡣࠊࣄࢺ TET3 㑇ఏᏊࡢ CXXC domainࠊCys-rich InsertࠊDSBHࠊLow-complexity insert ࢆඖ⾲♧ࡋ࡚࠸ࡿࠋྛ㡿ᇦࡢቃ⏺ࡣࠊ ࣑ࣀ㓟ࡢ␒ྕࡀグࡋ࡚࠶ࡿࠋࡲࡓࠊṇࡢ㑅ᢥࢆཷࡅࡓࢧࢺࢆڸ༳࡛࣑ࣀ㓟␒ྕඹ♧ࡋ࡚࠸ࡿࠋ PAML ゎᯒࡢ M8 ࣔࢹ᳨࡛ࣝฟࡉࢀࡓࢧࢺࡣ㟷Ⰽ࡛♧ࡍࠋ 3 – 5 ⣔⤫ἢࡗࡓࢧࢺ≉␗ⓗ㑅ᢥ㐣⛬ࡢኚ ␗࡞ࡿ㐍⣔⤫ἢࡗࡓࢥࢻࣥࢧࢺ≉␗ⓗ࡞⮬↛㑅ᢥࡢ㐣⛬ࢆ᫂ࡽࡍࡿࡓࡵࠊ᭱ ᑬἲࢆ⏝࠸ࡿFitmodel ࣉࣟࢢ࣒ࣛࡼࡿゎᯒࢆ⾜ࡗࡓࠋFitmodel ࣉࣟࢢ࣒ࣛࡣࠊࣈࣛࣥ ࢳࢧࢺࢥࢻࣥ⨨ࣔࢹࣝࢆ⏝ࡋ࡚࠸ࡿ [21, 22]ࠋࡇࡢࣔࢹࣝࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓ⣔⤫ ཬࡧࢥࢻࣥࢧࢺࡘ࠸࡚ࡢ๓ࡢሗࡀࡃ↓࠸≧ែ࡛ࠊࡍ࡚ࡢࣈࣛࣥࢳ㸦⣔⤫㸧ཬ ࡧࢥࢻࣥࢧࢺࢃࡓࡗ࡚ṇࡢ㑅ᢥࢆ᳨ฟྍ⬟࡞ࣔࢹ࡛ࣝ࠶ࡿ [42]ࠋFitmodel ࡛ᐇ⾜ྍ⬟ ࡞4 ✀ࡢࣔࢹࣝࠊM0ࠊM3ࠊM3 + S1ࠊM3 + S2 ࡛ࡢゎᯒ⤖ᯝࢆ⾲ 2 ⾲ 3 ♧ࡋࡓࠋ⾲ 2 ࡣࠊ3 ࡘࡢࣔࢹࣝ࣌㸦M0 vs. M3ࠊM3 vs. M3 + S1ࠊM3 + S1 vs. M3 + S2㸧ࡘ࠸࡚ࠊ ྛࣔࢹࣝࡢᑐᩘᑬᗘᕪࡢ2 ಸ㸦2ΔlnL㸧ࢆ♧ࡋ࡚࠸ࡿࠋP್ࡽ᫂ࡽ࡞ࡼ࠺ࠊTET1ࠊ TET2ࠊ TET3 ࡢ࠸ࡎࢀࡢ㑇ఏᏊ࡛ࡶࠊM3 + S2 ࣔࢹࣝࡀ᭱㐺࡛࠶ࡿࡇࡀศࡿࠋM3 + S2 ࡣࠊࢫࢵࢳࣥࢢࣔࢹࣝࡢ୰࡛ࡶ᭱ࡶᰂ㌾࡞ኚࢆチᐜࡍࡿࣔࢹ࡛ࣝ࠶ࡾࠊ3 ࡘࡢȰࢡࣛࢫ 㛫ࡢࢫࢵࢳࣥࢢࡘ࠸࡚ᆒ➼࡞㏿ᗘẚࢆ௬ᐃࡍࡿࣔࢹ࡛ࣝ࠶ࡿࠋ
19 ⾲
⾲2 Fitmodel ࣉࣟࢢ࣒ࣛࡼࡿྛࣔࢹࣝ࣌㛫ࡢᑬᗘẚ᳨ᐃ㸦LRTs㸧
M0 vs. M3 M3 vs. M3 + S1 M3 + S1 vs. M3 + S2
2ΔlnL P value 2ΔlnL P value 2ΔlnL P value
TET1 2858.77 P < 0.001 376.92 P < 0.001 71.11 P < 0.001 TET2 2803.21 P < 0.001 345.29 P < 0.001 63.69 P < 0.001 TET3 2926.57 P < 0.001 160.22 P < 0.001 12.59 P < 0.002 M0ࠊM3ࠊM3+S1ࠊM3+S2 ࡣࡑࢀࡒࢀࣔࢹࣝྡࢆ⾲ࡍࠋ ⾲3 ࡢ M3 + S2 ࣔࢹࣝࡢ⤖ᯝࡼࢀࡤࠊTET1 ࠾࠸࡚ࠊȰࡢ᥎ᐃ್ࡣȰ1 = 0.01ࠊȰ2 = 0.23ࠊȰ3 = 1.34 ࡛࠶ࡗࡓࠋȰ3ࡢ್ࡣ1 ࡼࡾࡶࡁࡃṇࡢ㑅ᢥࡢᏑᅾࢆ♧ࡋࠊࡑࡢྜ p3 ࡣ0.25 ࡛࠶ࡾࠊ㈇ࡢ㑅ᢥࢆ♧ࡍ 1 ᮍ‶ࡢȰ1ཬࡧȰ2ࡢྜࡢྜィࡣࠊ0.74 ࡛࠶ࡗࡓࠋࡍ࡞ ࢃࡕࠊTET1 㑇ఏᏊࡢࢥࢻࣥࢧࢺࡢ 25%ࡀ࠸ࡎࢀࡢ⣔⤫࡛ṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉ ࢀࡓࠋࡲࡓࠊȰ1Ȱ2ࡢࢫࢵࢳࣥࢢ࣮ࣞࢺࡀR12 = 0.71ࠊȰ㸯Ȱ3ࡢࢫࢵࢳࣥࢢ࣮ࣞ ࢺࡀR13 㸻 0.2 ࡛࠶ࡗࡓࡢᑐࡋ࡚ࠊȰ2Ȱ3ࡢࢫࢵࢳࣥࢢ࣮ࣞࢺࡣR23 = 3.34 ࡛࠶ࡾࠊ R23ࡀẚ್࡚ࡀࡁ࠸ࡇࡀศࡿࠋࡇࢀࡣࠊ┦ᑐⓗ㢮ఝࡋࡓ㑅ᢥࣃࢱ࣮ࣥ㛫㸦㢮 ఝࡋࡓȰ㸧࡛ࡢኚࡀⰋࡃ㉳ࡇࡗࡓࡇࢆ♧ࡋ࡚࠸ࡿࠋTET2 ࡘ࠸࡚ࡶࠊTET1 ྠᵝࡢ ഴྥࢆ♧ࡋࠊȰࡢ᥎ᐃ್ࡣȰ1 = 0.005ࠊȰ2 = 0.30ࠊȰ3 = 1.54ࠊp3 = 0.17ࠊp1ཬࡧp2ࡢ್ ࡢྜィࡣ0.83 ࡛࠶ࡗࡓࠋࡲࡓࠊࢫࢵࢳࣥࢢ࣮ࣞࢺ R23=4.34ࠊR12=0.74 ཬࡧ R13=0.33 ࡞ࡾࠊࡇࢀࡶࡲࡓ㢮ఝࡋࡓ㑅ᢥࣃࢱ࣮ࣥࡢኚࡀⰋࡃ㉳ࡇࡗࡓࡇࡀศࡿࠋࡇࢀࡽᑐ ࡋ࡚ࠊTET3 ࡛ࡣȰࡢ᥎ᐃ್ࡣȰ1 = 0.003ࠊȰ2 = 0.22ࠊȰ3 = 1.07 ࡛࠶ࡾࠊṇࡢ㑅ᢥࢆ♧ࡍ Ȱࡢ࢝ࢸࢦ࣮ࣜࡣᏑᅾࡋ࡞ࡗࡓࠋࡲࡓࠊȰ1 = 0.003 ࢆ♧ࡍࢥࢻࣥࡢྜࡣ p1 = 0.68 ࡞ ࡾࠊȰ2 = 0.22 ࡢྜ p2 = 0.23 ཬࡧȰ3 = 1.07 ࡢྜ p3 = 0.09 ࡢ⤖ᯝࡶྜࢃࡏ࡚ࠊTET3 ࡣయࡋ࡚ᙉ࠸㈇ࡢ㑅ᢥࢆཷࡅࡓ⪃࠼ࡽࢀࡿࠋ
20 ⾲ ⾲3 TET 㑇ఏᏊࡢ Fitmodel ࣉࣟࢢ࣒ࣛࡼࡿ᭱ᑬἲゎᯒ Ȱ1ࠊȰ2ࠊȰ3 ࡣࠊȰ1 <Ȱ2 <Ȱ3 ࡢࡁࡉࡢ㛵ಀ࡛᥎ᐃࡉࢀࡿ್࡛࠶ࡿࠋࡑࡢྜࡣࠊࡑࢀࡒࢀࠊp1ࠊp2ࠊ p3 ࡛♧ࡉࢀࡿࠋR ࡣࠊࡑࢀࡒࢀᩘᏐࡢ♧ࡍȰ㛫ࢆࢫࢵࢳࡍࡿ㏿ᗘ࡛࠶ࡿࠋ࠼ࡤࠊR12ࡣȰ1Ȱ2ࢆࢫ ࢵࢳࡍࡿ㏿ᗘ࡛࠶ࡿࠋ ḟࠊࢥࢻࣥࢧࢺࡀࡢ⛬ᗘࡢࣈࣛࣥࢳ(⣔⤫)࡛ṇࡢ㑅ᢥࢆཷࡅࡓࢆ᫂ࡽࡍࡿࡓ ࡵࠊࡑࢀࡒࢀࡢࢧࢺࡀṇࡢ㑅ᢥࢆཷࡅࡓࣈࣛࣥࢳᩘࢆホ౯ࡋࡓࠋAppendix 6 ♧ࡍ perl ࣉࣟࢢ࣒ࣛࢆ⏝ࡋ࡚ࠊࡑࢀࡒࢀࡢࢥࢻࣥࢧࢺࡢᚋ☜⋡PP ࡀ 0.9 ࡼࡾࡶࡁ࠸ࣈࣛࣥ ࢳࢆィᩘࡋࠊࡑࡢ⤖ᯝࢆᅗ10㹼ᅗ 12 ♧ࡍࠋTET1 ࡣ 2114 ಶࡢࢥࢻࣥࡽ࡞ࡾࠊࡑࡢ 14% ࡢ298 ಶࡢࢥࢻࣥࡣࠊ࠸ࡎࢀࡢࣈࣛࣥࢳ࡛Ȱ3 = 1.34 ࢆ♧ࡍṇࡢ㑅ᢥࡀ᳨ฟࡉࢀࡓ㸦ᅗ 10㸧ࠋTET2 ࡣ 1905 ಶࡢࢥࢻࣥࡽ࡞ࡾࠊࡑࡢ 6.7%ࡢ 128 ಶࡢࢥࢻ࡛ࣥȰ3 = 1.54 ࢆ♧ࡍ ṇࡢ㑅ᢥࡀ᳨ฟࡉࢀࡓ㸦ᅗ11㸧ࠋTET1ࠊTET2 ࡢ࠸ࡎࢀ࠾࠸࡚ࡶࠊ๓㏙ࡢࢧࢺࣔࢹࣝ ࡢ⤖ᯝࡢࡼ࠺ࠊCys-rich Insert 㡿ᇦࠊDSBH 㡿ᇦ࠾ࡼࡧ CXXC 㡿ᇦ࡛ࡣഹ࡞ࣈࣛࣥࢳ ࡛ࡋṇࡢ㑅ᢥࡀㄆࡵࡽࢀ࡞ࡗࡓࠋ୍᪉ࠊN ᮎ➃ഃࡢ༙ศࠊཬࡧ Low-complexity insert 㡿ᇦࡢከࡃࡢࢥࢻࣥࢧࢺ࡛ࡣࠊከᩘࡢࣈࣛࣥࢳ࡛ṇࡢ㑅ᢥࡀ᳨ฟࡉࢀࡓࠋTET3 ࡣࠊȰ3 = 1.07 ప࠸್࡛࠶ࡾࠊṇࡢ㑅ᢥ࠸࠺ࡼࡾࡣ㑅ᢥ⦆≧ែ࠸ࡗࡓ᪉ࡀⰋ࠸ࡀࠊ4.3%ࡢ 77 ಶࡢࢥࢻࣥࡀ࠸ࡎࢀࡢࣈࣛࣥࢳ࡛ࡇࡢȰ3 ࢆ♧ࡍࡇࡀ☜ㄆࡉࢀࡓࠋࡑࡋ࡚ࠊࡇࢀࡽ 77 ಶࡢࢥࢻࣥࡣࠊࡲࡓCys-rich Insert 㡿ᇦࠊDSBH ཬࡧ CXXC 㡿ᇦࡢእഃከࡃศᕸࡋ࡚ M0 M3 M3 + S1 M3 + S2 TET1: LnL -87135.73 -85706.34 -85517.88 -85482.33 ω1 ω2 ω3 0.39 0.04 0.35 0.90 0.00 0.42 1.21 0.01 0.23 1.34 p1 p2 p3 0.25 0.48 0.26 0.37 0.41 0.22 0.28 0.46 0.25 R12 R13 R23 1.54 1.54 1.54 0.71 0.2 3.34 TET2: LnL -75677.72 -74276.11 -74103.47 -74071.62 ω1 ω2 ω3 0.35 0.04 0.36 0.94 0.00 0.42 1.24 0.005 0.30 1.54 p1 p2 p3 0.31 0.50 0.19 0.39 0.44 0.17 0.35 0.48 0.17 R12 R13 R23 1.6 1.6 1.6 0.74 0.33 4.34 TET3: LnL -48719.65 -47256.37 -47176.26 -47169.96 ω1 ω2 ω3 0.11 0.01 0.19 0.75 0.00 0.28 0.98 0.003 0.22 1.07 p1 p2 p3 0.62 0.28 0.10 0.69 0.23 0.08 0.68 0.23 0.09 R12 R13 R23 2.14 2.14 2.14 1.04 2.27 9.96
21 ࠸ࡓ㸦ᅗ12㸧ࠋ ᅗ ᅗ10 TET1 㑇ఏᏊࢥࢻࣥࢧࢺ࡛ṇࡢ㑅ᢥࢆ♧ࡍࣈࣛࣥࢳᩘศᕸ TET1 㑇ఏᏊࡢ㓄ิ୰࡛ࠊȰ3 = 1.34 ࡀ᳨ฟࡉࢀࡿࣈࣛࣥࢳࡢᩘࢆࢥࢻࣥࡈ⾲♧ࡋ࡚࠸ࡿࠋᶓ㍈ࡣࢥࢻ ࣥ␒ྕࠊ⦪㍈ࡣPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࣈࣛࣥࢳࡢᩘࢆ⾲♧ࠋ ᅗ11 TET2 㑇ఏᏊࢥࢻࣥࢧࢺ࡛ṇࡢ㑅ᢥࢆ♧ࡍࣈࣛࣥࢳᩘศᕸ TET2 㑇ఏᏊࡢ㓄ิ୰࡛ࠊȰ3 = 1.54 ࡀ᳨ฟࡉࢀࡿࣈࣛࣥࢳࡢᩘࢆࢥࢻࣥࡈ⾲♧ࡋ࡚࠸ࡿࠋᶓ㍈ࡣࢥࢻ ࣥ␒ྕࠊ⦪㍈ࡣPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࣈࣛࣥࢳࡢᩘࢆ⾲♧ࠋ
22 ᅗ ᅗ12 TET3 㑇ఏᏊࢥࢻࣥࢧࢺ࡛㑅ᢥ⦆ࢆ♧ࡍࣈࣛࣥࢳᩘศᕸ TET3 㑇ఏᏊࡢ㓄ิ୰࡛ࠊȰ3 = 1.07 ࡀ᳨ฟࡉࢀࡿࣈࣛࣥࢳࡢᩘࢆࢥࢻࣥࡈ⾲♧ࡋ࡚࠸ࡿࠋᶓ㍈ࡣࢥࢻ ࣥ␒ྕࠊ⦪㍈ࡣPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࣈࣛࣥࢳࡢᩘࢆ⾲♧ࠋ ᅗ 13 ࡣࠊಶࠎࡢࣈࣛࣥࢳ࠾࠸࡚Ȱ3ࢆ♧ࡍࢥࢻࣥࢧࢺᩘࡑࡢࣈࣛࣥࢳᩘࡢศᕸࢆ ⾲♧ࡋ࡚࠸ࡿࠋྛࣈࣛࣥࢳ࡛Ȱ3ࢆ♧ࡍࢥࢻࣥࢧࢺᩘࡣࠊAppendix 8 ᥖ㍕ࡋࡓ⊂⮬ సᡂࡋࡓPerl ࣉࣟࢢ࣒࡛ࣛ㞟ィࡋࡓࠋᅗࡽศࡿࡼ࠺ࠊྛࣈࣛࣥࢳ࠾࠸࡚Ȱ3ࢆ♧ࡍ
ࢥࢻࣥࢧࢺࡢᩘࡣࠊTET1 ࡛ࡣ 0 ࡽ 168ࠊTET2 ࡛ࡣ 0 ࡽ 107ࠊTET3 ࡛ࡣ 0 ࡽ 64
ࡉࡲࡊࡲ࡛࠶ࡗࡓࠋȰ3ࡢࢥࢻࣥࢧࢺࡀከᩘᏑᅾࡍࡿࣈࣛࣥࢳࡣ㐍㏿ᗘࡀࡁ࠸࠸ ࠼ࡿࡢ࡛ࠊḟࠊTET 㑇ఏᏊࡢ⣔⤫ᶞ୰࡛ࡢ࠶ࡓࡾࡢࣈࣛࣥࢳȰ3ࡢࢥࢻࣥࢧࢺࡀከ ࠸ࢆ᳨ウࡋࡓࠋᅗ4ࠊᅗ 5ࠊᅗ 6 ࡢ⣔⤫ᶞ୰ࡑࢀࡒࢀ㟷ࠊ࢜ࣞࣥࢪࠊ⥳࡛⾲♧ࡋࡓࡼ࠺ ࠊTET1ࠊTET2ࠊTET3 㑇ఏᏊࡢ࠸ࡎࢀࡢሙྜࡶ⣔⤫ᶞࡢ῝㒊ࡢࣈࣛࣥࢳ㞟୰ࡋࡓࠋ ࡇࡢࡇࡣࠊ့ங㢮ࡢ♽ඛࡀศᒱከᵝࡍࡿᮇᑐᛂࡋ࡚TET 㑇ఏᏊࡢ㐍㏿ᗘࡀቑ ࡋ࡚࠸ࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋ
23 ᅗ ᅗ13 TET 㑇ఏᏊ㓄ิࡢȰ3ࡢ್ࢆ♧ࡍࢧࢺᩘࡑࡢࣈࣛࣥࢳᩘ ᶓ㍈ࡣࠊPP > 0.9 ࡛Ȱ3ࡢ್ࢆ♧ࡍࢧࢺࡢᩘࠊ⦪㍈ࡣࠊࡑࡢࢧࢺᩘࡀ᳨ฟࡉࢀࡓࣈࣛࣥࢳࡢᩘࢆ⾲♧ࠋ 3 - 6 3 ḟඖᵓ㐀࣐ࢵࣆࣥࢢ ṇࡢ㑅ᢥࡢᯝࡓࡍᙺࡢゎ᫂ࡢࡓࡵࠊTET ࢱࣥࣃࢡ㉁ࡢᵓ㐀୰ࡢࡇṇࡢ㑅ᢥࢆཷࡅ ࡓ࣑ࣀ㓟ࡀ⨨ࡍࡿࡢࢆࠊࡑࡢ 3 ḟඖࣔࢹࣝୖ࣐ࢵࣆࣥࢢࢆ⾜ࡗ࡚☜ࡵࡓࠋTET ࢱࣥࣃࢡ㉁ࡢ❧యᵓ㐀ࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿࡢࡣ TET2 ࡢゐ፹ࢥࢻ࣓ࣥࡢࡳ࡛࠶ࡿࡓ ࡵࠊᮏ◊✲࡛ࡣTET2 ࡢࡳࡘ࠸࡚࣐ࢵࣆࣥࢢࢆ⾜ࡗࡓࠋᅗ 14 ࣐ࢵࣆࣥࢢࢆ⾜ࡗࡓ TET2
ࡢゐ፹㡿ᇦ㸦PDB ID:4NM6㸧ࢆ♧ࡍࡀࠊࡇࢀࡣࣄࢺࡢ TET2 ࡀ DNA ⤖ྜࡋࡓ≧ែࡢ⤖ ᬗᵓ㐀ᇶ࡙࠸ࡓࡶࡢ࡛࠶ࡿ [13]ࠋ
TET2 ࡢゐ፹ࢥࢻ࣓࡛ࣥṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀࡓ࣑ࣀ㓟ࢧࢺࡣࠊPAML
ཬࡧDatamonkey ࡼࡗ᳨࡚ฟࡉࢀࡓ㸯ࢧࢺ㸦1312L㸧ࠊFitmodel ࡢゎᯒࡼࡾ᳨ฟ
ࡉࢀࡓ4 ࢧࢺ㸦1203S, 1204S, 1205S, 1892N㸧ࡢྜィ 5 ࡘ࡛࠶ࡿࠋࡇࢀࡽࡢࢥࢻࣥࢧ
24 ᢥࡀ᳨ฟࡉࢀࡓ5 ࡘࡢ࣑ࣀ㓟ࢧࢺࡣࠊ4 ࡘࡣゐ፹ࢥࢻ࣓ࣥࡢ Cys-rich Insert 㡿ᇦ ࠊ1 ࡘࡣ DSBH 㡿ᇦᏑᅾࡋࡓࠋศᏊ⾲㠃ࡢ⾲♧ࢆ⾜ࡗࡓࡶࡢࡀᅗ 14(B)࡛࠶ࡿࡀࠊṇ ࡢ㑅ᢥࢆཷࡅࡓ5 ࡘࡢ࣑ࣀ㓟ࢧࢺࡣࠊ࡚ࢱࣥࣃࢡ㉁⾲㠃⨨ࡋ࡚࠸ࡓࠋ ᅗ ᅗ14 ࣄࢺ 7(72 ࡢゐ፹㡿ᇦࡢ 3 ḟඖᵓ㐀
㸦A㸧: ⥳Ⰽࣜ࣎ࣥࡣࠊDNA 㔜㙐ࡢ」ྜయ࡞ࡗࡓࣄࢺ TET2 ࡢゐ፹㡿ᇦࢆ⾲ࡍࠋ㯤Ⰽ㒊ศࡣ DNA ࢆ♧ࡍࠋ࣐ࢮࣥࢱⰍ㒊ศࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓ5 ࡘࡢࢧࢺࡢ࣑ࣀ㓟ഃ㙐ࢆ♧ࡍࠋ㸦B㸧: ࣄࢺ TET2 ࡢ ゐ፹㡿ᇦࡢศᏊ⾲㠃ࢆ⾲ࡍࠋ㟷Ⰽ㒊ศࡣࠊṇࡢ㑅ᢥࢆཷࡅࡓ5 ࡘࡢࢧࢺࡢ࣑ࣀ㓟ഃ㙐ࢆ♧ࡍࡀࠊ࡚ ⾲㠃⨨ࡍࡿ㸦Akahori et al., 2015 [48]ࡼࡾࠊ୍㒊ᨵኚࠊ㌿㍕㸧ࠋ 3 - 7 ࣑ࣀ㓟ࡢᩡᛶ⨨࠾ࡼࡧศᒱᛶ⨨ codeMLancestral ࣉࣟࢢ࣒ࣛ [38]ࢆ⏝ࡋࠊTET 㑇ఏᏊ⣔⤫ᶞࡢࣈࣛࣥࢳẖࡢẚ㍑ᇶ ࡙࠸࡚ࠊᩡᛶ࣑ࣀ㓟⨨ศᒱᛶ࣑ࣀ㓟⨨ࢆゎᯒࡋࡓࠋゎᯒ⤖ᯝࡢ㞟ィࡣࠊ Appendix 7 ♧ࡍ⊂⮬సᡂࡋࡓ Perl ࣉࣟࢢ࣒ࣛࢆ⏝ࡋࡓࠋᅗ 15 ࡣࠊTET1ࠊTET2
ཬࡧ TET3 ࡘ࠸࡚ࠊ࡚ࡢࣈࣛࣥࢳ࣌ࡢᩡᛶ⨨ࡢᚋ☜⋡ࡢࠊศᒱᛶ⨨ࡢ
ᚋ☜⋡ࡢࢆࣉࣟࢵࢺࡋ࡚♧ࡋࡓࠋࡢ◊✲⤖ᯝ [38, 43]ྠᵝࠊᩡᛶ⨨ศᒱᛶ ⨨ࡢ㢖ᗘࡢ㛫ࡣ┤⥺㛵ಀࡀㄆࡵࡽࢀࠊศᒱᛶ⨨ࡢ᪉ࡀᩡᛶ⨨ࡼࡾࡶ㢖ᗘࡀ㧗࠸ࠋ
25
┦ᑐⓗศᒱᛶ⨨ࡀᑡ࡞ࡃࠊᩡᛶ⨨ࡀከ࠸ഴྥࡀ♧ࡉࢀࡓࠋࡇࡢࡇࡣࠊ့ங㢮ࡢ 㐍࠾࠸࡚ࠊTET3 ࡣ TET1 ࠾ࡼࡧ TET2 ࡣࡁࡃ␗࡞ࡗࡓ㐍ືែࢆ⤒࡚ࡁࡓࡇ ࢆ♧၀ࡋ࡚࠸ࡿࠋ
ᅗ
ᅗ15 TET 㑇ఏᏊࡢᩡᛶ⨨࠾ࡼࡧศᒱᛶ⨨
ྛTET 㑇ఏᏊ⣔⤫ᶞ࠾࠸࡚ࡢࠊ࡚ࡢࣈࣛࣥࢳ࡛࣌ࡢᩡᛶ⨨ࡢᚋ☜⋡࠾ࡼࡧศᒱᛶ⨨ࡢᚋ ☜⋡ࡢྜィࢆࡑࢀࡒࢀࣉࣟࢵࢺࡋࡓࠋTET1 ࢆ㟷ⰍࠊTET2 ࢆ⥳ⰍࠊTET3 ࢆ㉥Ⰽ࡛♧ࡍࠋ
26
44. ⪃ᐹ
TET ࣇ࣑࣮ࣜ㑇ఏᏊࡣࠊ⣽⬊ࡢศࡸⓎ⏕ࡢᵝࠎ࡞ᒁ㠃ࡢไᚚ㛵ಀࡋ࡚࠸ࡿࠋࡇࡢ ࡇࢆᫎࡋ࡚ࠊ⣽⬊ࡢࢱࣉࡸྛ⤌⧊࡛ࡢTET ࣇ࣑࣮ࣜ㑇ఏᏊࡢⓎ⌧ࣞ࣋ࣝࡣࡁࡃ ኚືࡋࠊࡲࡓࣃࣛࣟࢢ㛫࡛ࡶⓎ⌧ࡀࡁࡃ␗࡞ࡗ࡚࠸ࡿ [26, 44, 45]ࠋࡓ࠼ࡤࠊ࣐࢘ࢫࡢ ES ⣽⬊࡛ࡣ TET1ࠊTET2 ࡢⓎ⌧ࡀⴭࡋࡃ㧗࠸ࡀࠊTET3 ࡘ࠸࡚ࡣ࣐࢘ࢫࡢ༸ẕ⣽⬊ࡸ ึᮇࡢཷ⢭༸࡛Ⓨ⌧ࡀୖ᪼ࡍࡿ㸦ᅗ16㸧ࠋཷ⢭┤ᚋࡢ⬇Ⓨ⏕ึᮇẁ㝵࠾࠸࡚ࠊ㞝ᛶ๓᰾࡛ ࡣTET3 ፹ࡉࢀࡓ DNA ࢩࢺࢩࣥሷᇶ㞟ᅋࡢᗈ⠊࡞㓟ࡀ㉳ࡇࡿ [45]ࠋࡲࡓࠊ⬇┙⬊ ᮇࡢⓎ⏕ẁ㝵࡛ࡣࠊES ⣽⬊࡞ࡿෆ㒊⣽⬊ሢ࡛つᶍ࡞ DNA ࡢ࣓ࢳࣝࡀㄏⓎࡉࢀࡿࠋ ࡇࡢⓎ⏕ẁ㝵࡛ࡶTET1ࠊTET2 ࡣ㧗ᗘⓎ⌧ࡋࠊ࠾ࡑࡽࡃ DNA ࣓ࢳࣝࣃࢱ࣮ࣥࡢᵓ ⠏࠾ࡼࡧㄪᩚࢆ⾜ࡗ࡚࠸ࡿ [7]ࠋࡇࢀࡽࡢ TET ࣇ࣑࣮ࣜ㑇ఏᏊ㛫ࡢ␗࡞ࡗࡓⓎ⌧ࣃࢱ࣮ ࣥࡣࠊⓎ⏕ศ㐣⛬࠾ࡼࡧᵝࠎ࡞⤌⧊࠾࠸࡚ࠊTET 㑇ఏᏊࡀಶูࡢ㔜せ࡞⏕≀Ꮫⓗᶵ⬟ ࢆᢸࡗ࡚࠸ࡿࡇࢆ♧ࡋ࡚࠸ࡿ [45]ࠋᮏ◊✲࡛ࡣࠊࡇ࠺ࡋࡓ TET 㑇ఏᏊࡢᶵ⬟ศ㛵ࢃ ࡿ㐍ⓗせᅉࠊࡾࢃࡅ⮬↛㑅ᢥࡢᙺࢆ᫂ࡽࡍࡿࡓࡵࠊTET ࣇ࣑࣮ࣜ㑇ఏᏊࡢ ศᏊ㐍ゎᯒࢆ⾜ࡗࡓࠋ ᅗ16 TET 㑇ఏᏊࡢⓎ⌧ࣞ࣋ࣝ TET 㑇ఏᏊࡢయࡋ࡚ࡢศᏊ㐍ࡢ≉ᚩࡣࠊPAML ࣉࣟࢢ࣒ࣛࡢ M0 ࣔࢹࣝࡽ᥎ᐃ ࡉࢀࡓࠋࡍ࡞ࢃࡕࠊM0 ࣔࢹࣝࡼࡾᚓࡽࢀࡿྠ⩏⨨㏿ᗘᑐࡍࡿ㠀ྠ⩏⨨㏿ᗘࡢẚ࡛ ࠶ࡿȰ್ᇶ࡙࠸࡚ࠊ့ங㢮ࡢTET 㑇ఏᏊ࡛ࡣ㈇ࡢ㑅ᢥࡀᨭ㓄ⓗ࡛࠶ࡿࡇࡀ᫂ࡽ࡞ ࡗࡓࠋࡇࡢ⤖ᯝࡣࠊTET 㑇ఏᏊࡀయࡋ࡚ࡣಖᏑⓗ⥔ᣢࡉࢀ࡚࠸ࡿࡇࢆ♧ࡋ࡚࠾ࡾࠊ ့ங㢮ࡢ⏕⌮ᶵ⬟ࡸ⏕Ꮡ࠾ࡅࡿTET 㑇ఏᏊᶵ⬟ࡢ㔜せࡉࢆ♧၀ࡋ࡚࠸ࡿ [46]ࠋࡲࡓࠊࢥ ࢻࣥࢧࢺ࡛ࣞ࣋ࣝࡢ㑅ᢥࣃࢱ࣮ࣥࡢኚࢆ≉ᚩࡅࡿࢧࢺࣔࢹࣝࢆ⏝࠸࡚ࠊṇࡢ㑅ᢥ ࢆ♧ࡍ࠸ࡃࡘࡢࢥࢻࣥࢧࢺࡢᏑᅾࡀ᫂ࡽ࡞ࡗࡓࠋḟࠊࡇࢀࡽṇࡢ㑅ᢥࡀ࡚ࡢ TET ࣇ࣑࣮ࣜ㑇ఏᏊྠᵝാࡃࡢྰࢆゎᯒࡋࡓࠋPAML ࣉࣟࢢ࣒ࣛࡢ M8 ࣔࢹࣝ27 ࡼࢀࡤࠊTET1 ࠾ࡼࡧ TET2 ࡣ୍㒊ࡢࢥࢻࣥࢧࢺ࡛ṇࡢ㑅ᢥࢆཷࡅࡓࡇࡀศࡗࡓࡀࠊ TET3 ࡛ࡣṇࡢ㑅ᢥࢆ࠸ࡎࢀࡢࢧࢺ࡛ࡶぢ࠸ࡔࡍࡇࡀฟ᮶࡞ࡗࡓࠋ Fitmodel ࣉࣟࢢ࣒࡛ࣛࡣࠊTET ࣇ࣑࣮ࣜ㑇ఏᏊ⣔⤫ᶞࡢᵝࠎ࡞ࣈࣛࣥࢳ࡛ࡢࢧࢺ≉ ␗ⓗ࡞㑅ᢥࣃࢱ࣮ࣥࡢ㐪࠸ࢆᗄࡘࡢࣃ࣓࣮ࣛࢱࢆ⏝࠸࡚ㄪࡿࡇࡀ࡛ࡁࡓ [21, 42]ࠋ Fitmodel ࡼࡗ࡚ᚓࡽࢀࡓ⤖ᯝࡣࠊᇶᮏⓗࡣࢧࢺࣔࢹࣝࢆ⏝ࡋࡓ PAML ゎᯒࡢ⤖ᯝ ఝ࡚࠾ࡾࠊTET 㑇ఏᏊࡢ㐍ࡢ㒊ศ࠾࠸࡚㈇ࡢ㑅ᢥࡀ㔜せ࡞ᙺࢆᯝࡓࡋ࡚ࡁࡓࡇ ࢆ♧ࡋ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊᮏ◊✲࡛ヲ⣽ゎᯒࡋࡓࣈࣛࣥࢳࢧࢺ≉␗ⓗ࡞㑅ᢥ ࣃࢱ࣮ࣥࡢኚࡣࠊTET ࣇ࣑࣮ࣜ㑇ఏᏊࡢࡑࢀࡒࢀ࡛ࡁࡃ␗࡞ࡗ࡚࠸ࡓࠋTET1 ࠾ࡼ ࡧ TET2 㑇ఏᏊ࡛ࡣࠊ㐍ࡢ㐣⛬࡛」ᩘࡢࣈࣛࣥࢳἢࡗ࡚ṇࡢ㑅ᢥࢆཷࡅࡓከࡃࡢࢥࢻ ࣥࢧࢺࡀ♧ࡉࢀࡓࠋࡇࢀࡣࠊ့ங㢮ࡀከᵝࡍࡿ㐣⛬࡛TET1 ࠾ࡼࡧ TET2 㑇ఏᏊࡀᛴ ㏿㐍ࡋࡓࡇࢆ♧ࡋ࡚࠾ࡾࠊ࠾ࡑࡽࡃࡣࡑࡢ⤖ᯝࡋ࡚㑇ఏᏊࡢᶵ⬟ศ࣭ከᵝࡀ ㉳ࡇࡗࡓ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽࠊTET3 㑇ఏᏊ࡛ࡣࠊPAML ࣉࣟࢢ࣒ࣛࡲࡓࡣ Fitmodel ࣉࣟࢢ࣒ࣛࡢఱࢀࡼࡗ࡚ࡶࠊṇࡢ㑅ᢥࢆཷࡅࡓ࠸࠺༑ศ࡞ドᣐࡀᚓࡽࢀ࡞ ࡗࡓࠋࡇ࠺ࡋࡓ⤖ᯝࡽࠊTET ࣇ࣑࣮ࣜ㑇ఏᏊࡢ᰿ᗏ࠶ࡿ㐍࣮ࣔࢻࡣࠊḟࡢࡼ࠺ ≉ᚩࡅࡿࡇࡀ࡛ࡁࡿࠋࡍ࡞ࢃࡕࠊTET3 ࡣࡑࡢ㓄ิࡀẚ㍑ⓗಖᏑࡉࢀ࡚࠾ࡾࠊTET1 ࠾ ࡼࡧ TET2 ࡣࡼࡾኚࡋ࡚ࡁࡓࠋᩡᛶࡢ࣑ࣀ㓟⨨ࡢゎᯒࡼࡗ࡚ᚓࡽࢀࡓ⤖ᯝ㸦ᅗ 15㸧ࡶࡲࡓࠊTET ࣇ࣑࣮ࣜ㑇ఏᏊࡢ␗࡞ࡗࡓ㐍ࢲࢼ࣑ࢡࢫࢆࡅࡿࡶࡢ࡛࠶ࡿࠋ TET3 㑇ఏᏊࡀኚࡋ࡚࠸࡞࠸⌮⏤ࡣࠊ㈇ࡢ㑅ᢥࡀാ࠸ࡓࡇࡼࡿ⪃࠼ࡽࢀࡿࠋ ࡇࡢࡇࡣࡲࡓࠊTET3 ࡢᶵ⬟ࡀ TET1ࠊTET2 ࡼࡾ௨ୖ⏕Ꮡᚲ㡲࡛࠶ࡿࡇࢆ♧၀ࡋ ࡚࠸ࡿࠋ⌧Ⅼ࡛▱ࡽࢀ࡚࠸ࡿ TET3 ࡢ᭱ࡶ㢧ⴭ࡞ᶵ⬟ࡣࠊཷ⢭┤ᚋࡢ㞝ᛶ๓᰾㸦⢭Ꮚ⏤ ᮶᰾㸧Ꮡᅾࡍࡿ DNA ࢩࢺࢩࣥሷᇶࡢ㓟ⓗ⬺࣓ࢳ࡛ࣝ࠶ࡿ [45]ࠋࡇࢀࡼࡾࠊ⢭Ꮚ ⏤᮶㑇ఏሗࡢⓎ⌧ࡀึࡵ࡚ྍ⬟࡞ࡿࡇࡽࠊಶయⓎ⏕ࡢึᮇࢫࢵࢳࡋ࡚ TET3 ࡢᏑᅾࡣྍḞ࡛࠶ࡿࠋࡋࡓࡀࡗ࡚ࠊTET3 ࡣᚋ⏕ື≀࠾ࡅࡿⓗ☜࡞᭷ᛶ⏕Ṫ࣭Ⓨ⏕ࢆಖ ドࡍࡿࡋࡃࡳࡢ୍㒊ࡋ࡚ࠊ⏕Ꮡᚲ㡲ࡢᙺࢆᢸࡗ࡚࠸ࡿ᥎ᐃࡉࢀࡿࠋࡲࡓࠊ᭷ᛶ⏕ Ṫࡢ㉳※ࡀᚋ⏕ື≀ࡢ㉳※௨ୖྂ࠸ࡇࢆ⪃៖ࡍࢀࡤࠊTET3 ࡣ TET ࣇ࣑࣮ࣜ㑇ఏᏊ ࡢ୰࡛ࡣࡑࡢ㉳※ࡀ᭱ࡶྂ࠸㑇ఏᏊ࡛࠶ࡿྍ⬟ᛶࡀ࠶ࡿࠋ TET ࢱࣥࣃࢡ㉁ࡢゐ፹ࢻ࣓ࣥࡣ࢝ࣝ࣎࢟ࢩࣝᮎ➃࠶ࡾࠊDNA ࡢ CpG 㓄ิࢆㄆ㆑ࡋ ࡚ 5-࣓ࢳࣝࢩࢺࢩࣥࡢ㓝⣲ⓗ㓟ࢆ⾜࠺ࠋࡇࡢゐ፹ࢻ࣓ࣥࡘ࠸࡚ࡢᵓ㐀࠾ࡼࡧኚ␗ᑟ ධᐇ㦂ࡢ◊✲ [8, 13]ࡽࠊ㓝⣲άᛶ࠾ࡼࡧ⿵ຓᅉᏊࡢ㓄ྥᛶᚲ㡲ࡢ࣑ࣀ㓟ṧᇶࡣ᪤ ᫂ࡽࡉࢀ࡚࠸ࡿࠋᮏ◊✲࡛ࡣࠊTET2 ࡢゐ፹ࢻ࣓࡛ࣥṇࡢ㑅ᢥࢆཷࡅࡓ 5 ࡘࡢ࣑ࣀ 㓟ࢧࢺࢆྠᐃࡋࡓࡀࠊࡇࢀࡽࡢࢧࢺࡣゐ፹άᛶỴᐃⓗ࡞ࡿ࣑ࣀ㓟ࡸࠊ⿵ຓᅉᏊ
28 ࡢ㓄ྥ㛵ࢃࡿ࣑ࣀ㓟ࡣ୍⮴ࡋ࡞ࡗࡓࠋࡇࡢࡇࡣࠊࡇࢀࡽゐ፹άᛶ㔜せ࡞࣑ ࣀ㓟ṧᇶࡣࠊࡑࡢ㔜せᛶࡢࡓࡵኚࢆチᐜࡉࢀࡎࠊ㐍ⓗᙉࡃಖᏑࡉࢀ࡚࠸ࡿࡇࢆ ♧ࡋ࡚࠸ࡿࠋ୍᪉ࠊ⯆῝࠸ࡇࠊゐ፹ࢻ࣓ࣥࡢ୰࡛ṇࡢ㑅ᢥࡀ᳨ฟࡉࢀࡓ࡚ࡢ ࣑ࣀ㓟ࢧࢺࡣࠊࢱࣥࣃࢡ㉁ࡢእഃ⾲㠃Ꮡᅾࡋ࡚࠸ࡓࠋ㏻ᖖࠊࢱࣥࣃࢡ㉁⾲㠃⨨ࡍ ࡿ࣑ࣀ㓟ࡣࠊࡢࢱࣥࣃࢡ㉁ࡢ┦స⏝㛵࡛ࡁࡿࠋࡋࡓࡀࡗ࡚ࠊṇࡢ㑅ᢥࢆཷࡅ ࡓ࣑ࣀ㓟ࡣࠊ㓝⣲⾲㠃࡛ࡢࢱࣥࣃࢡ㉁㛫┦స⏝ࢆࡋ࡚ࠊ㓝⣲ไᚚࢩࢫࢸ࣒ࡢᶵ⬟ศ ᙳ 㡪 ࡋ ࡚ ࠸ ࡿ ྍ ⬟ ᛶ ࡀ ᙉ ࡃ ♧ ၀ ࡉ ࢀ ࡿ ࠋBioGRID ࢹ ࣮ ࢱ ࣋ ࣮ ࢫ (https:// thebiogrid.org/)ࢆ⏝࠸࡚ࠊTET2 ┦స⏝ࡍࡿࢱࣥࣃࢡ㉁ࡘ࠸᳨࡚⣴ࡍࡿࠊࣄࢺ࣐ ࢘ࢫࡢ㉁㔞ศᯒἲ࡞ࡼࡿ⥙⨶ⓗゎᯒࡽࠊVPRBPࠊTBC1D4ࠊTCF3ࠊPAN2ࠊOGTࠊ NONOࠊSEMG1ࠊDDB1ࠊSAP30ࠊSUDS3ࠊSAP130ࠊHDAC2ࠊRBBP7ࠊHDAC1ࠊARID4Bࠊ SIN3A ࡞ࡢࢱࣥࣃࢡ㉁ࡀᐇ㝿⤖ྜࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡓࠋࡇࢀࡽࡢࢱࣥࣃࢡ㉁ ࡘ࠸࡚ࠊࢪ࣮ࣥ࢜ࣥࢺࣟࢪ࣮ゎᯒࢆ⾜࠺ࠊ㑇ఏᏊⓎ⌧ࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡไᚚࠊࣄࢫ ࢺࣥࡢ⬺ࢭࢳࣝ㓝⣲HDACࠊࢭࢳࣝࡼࡿ TP53 㑇ఏᏊάᛶࡢไᚚࠊ࡞ࡢ࢚ࣆ ࢪ࢙ࢿࢸࢵࢡไᚚ㛵ࢃࡿࢱࣥࣃࢡ㉁ࡀከࡃྵࡲࢀ࡚࠸ࡿࡇࡀศࡗࡓ㸦ᅗ17㸧ࠋ ᅗ17 TET2 ┦స⏝ࢱࣥࣃࢡ㉁ࡢࢪ࣮ࣥ࢜ࣥࢺࣟࢪ࣮ゎᯒ⤖ᯝ ᶓ㍈ࡣࠊྛࢱ࣮࣒࠾ࡅࡿ㑇ఏᏊࡢྜࢆ♧ࡋࠊࢢࣛࣇࡢᲬࡢඛࡢ್ࡣࠊ㑇ఏᏊᩘࢆ⾲ࡍࠋ ୍᪉ࠊUniProt ࢹ࣮ࢱ࣮࣋ࢫࡽࣄࢺయ⣽⬊✺↛ኚ␗ࡢሗࢆྲྀᚓࡋࠊᮏ◊✲᳨࡛ฟࡋࡓ ṇࡢ㑅ᢥࢆཷࡅࡓࢥࢻࣥࢧࢺẚ㍑ࡍࡿࠊࢧࢺࡢ࠸ࡃࡘࡣ⒴㛵㐃ࡍࡿ✺↛ኚ␗ ྠࡌࢧࢺ࠶ࡿ࠸ࡣࡑࡢ㏆ഐ⨨ࡍࡿࡇࡀศࡗࡓࠋ࠼ࡤࠊTET2 ࡢࢥࢻࣥࢧࢺ 123ࠊ817ࠊ1204ࠊ762ࠊ1811 ࡣ Fitmodel ࣉࣟࢢ࣒ࣛࡼࡾṇࡢ㑅ᢥࢆཷࡅࡓ᥎ᐃࡉࢀ ࡓࡀࠊࡇࢀࡽࡣ㦵㧊ቑṪᛶࡢ⭘⒆ࡳࡽࢀࡿ✺↛ኚ␗ࡢࢥࢻࣥࢧࢺ୍⮴ࡋࡓ [47]ࠋࡋ ࡋ࡞ࡀࡽࠊࡇࢀࡽࡢࢥࢻࣥࢧࢺࡢ⭘⒆ࡢ㛵ࡢ⛬ᗘࡣ⌧ᅾࡢࡇࢁ࡛᫂࠶ࡾࠊTET ࢱࣥࣃࢡ㉁୰࡛ࡢṇࡢ㑅ᢥࡢព⩏ࡶྵࡵ࡚ᚋࡢࡉࡽ࡞ࡿ◊✲ࡀᚲせ࡛࠶ࡿࠋ
29
55. ⥲ᣓ
ᮏ◊✲࡛ࡣࠊ့ங㢮ࡢᗈ⠊࡞TET 㑇ఏᏊࣇ࣑࣮࣓ࣜࣥࣂ࣮ࢆྠᐃࡍࡿࡶࠊศᏊ 㐍ゎᯒࡼࡗ࡚ࠊTET1ࠊTET2ࠊTET3 ྛࣃࣛࣟࢢ㛫ࡢ㐍Ꮫⓗ≉ᛶࢆ᫂ࡽࡋࡓࠋ ࡲࡓࠊࡇࢀࡽࡢ㑇ఏᏊ࡛ാࡃඃ࡞㑅ᢥᅽࡣ㈇ࡢ㑅ᢥ࡛࠶ࡗࡓࡀࠊ≉ᐃࡢ࣑ࣀ㓟ࢧࢺ ࡛ࡣᶵ⬟ⓗศ࠾ࡼࡧ㐺ᛂ㐍ࡢೃࢆ♧ࡍṇࡢ㑅ᢥࢆ☜ㄆࡋࡓࠋTET3 㑇ఏᏊẚࠊ TET1 ࠾ࡼࡧ TET2 㑇ఏᏊ࡛ࡣ㢖⦾ṇࡢ㑅ᢥࢆཷࡅࡓࡇࢆ᫂ࡽࡋࠊTET3 㑇ఏᏊࡀⴭࡋࡃಖᏑࡉࢀ࡚㐍ࡋ࡚ࡁࡓࡇࢆ♧ࡋࡓࠋTET2 ゐ፹ࢥࢻ࣓࡛ࣥࡣࠊ5 ࡘࡢṇࡢ㑅
ᢥࢆཷࡅࡓࢧࢺࡀࢱࣥࣃࢡ㉁⾲㠃ぢ࠸ࡔࡉࢀࡓࡀࠊࡇࢀࡽࡢࢧࢺࡀ㑇ఏᏊࡢゐ፹≉ ᛶࢆኚࡉࡏ࡚࠸ࡿྍ⬟ᛶࡶ᥎ᐹࡉࢀࡿࠋᚋࠊࡇ࠺ࡋࡓศᏊ㐍ゎᯒࡢ⤖ᯝᇶ࡙࠸࡚ ࡉࡽ࡞ࡿ◊✲ࢆ㐍ࡵࡿࡇࡼࡾࠊTET 㑇ఏᏊࣇ࣑࣮ࣜࡢࡼࡾヲ⣽࡞ᐇయࡢゎ᫂ࡀᮇᚅ ࡛ࡁࡿࠋ
30
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34
ㅰ
ㅰ ㎡
ᮏ◊✲ࡢ㐙⾜ཬࡧㄽᩥࡢసᡂ࠶ࡓࡾࠊከ࡞ࡈᣦᑟࢆ㈷ࡾࡲࡋࡓᒱ㜧ᏛᏛ㝔㐃ྜ ⸆་⒪ሗ◊✲⛉ Ṋ⸨ྜྷᚨ ᩍᤵᚰࡼࡾឤㅰ࠸ࡓࡋࡲࡍࠋ ࡲࡓࠊᮏㄽᩥࢆࡈ⢭ㄞ࠸ࡓࡔࡁࠊከࡃࡢࡈຓゝࢆ㈷ࡾࡲࡋࡓᒱ㜧ᏛᏛ㝔㐃ྜ⸆་ ⒪ሗ◊✲⛉ ຍ⸨ၿ୍㑻 ᩍᤵࠊୖ⏣ᾈ ᩍᤵ῝ㅰ࠸ࡓࡋࡲࡍࠋ ◊✲ࢆ㐍ࡵࡿ࠶ࡓࡾࠊᮏ◊✲㡿ᇦࡢࢆ࠶ࡲࡾ⌮ゎࡋ࡚࠸࡞࠸⚾㆟ㄽࢆࡋ࡚ࠊ⌮ ゎࢆᡭຓࡅࡋ࡚࠸ࡓࡔ࠸ࡓྜྷᓮ⣧ኵࡉࢇឤㅰ࠸ࡓࡋࡲࡍࠋᮏ◊✲ࡣࠊUniversity of Auckland ࡢ Dr. Stéphane Guindon Ặࡢసᡂࡉࢀࡓࣉࣟࢢ࣒ࣛࢆ⏝ࡋ࡚࠸ࡲࡍࠋࡇࡇ ཌࡃ࠾♩⏦ࡋୖࡆࡲࡍࠋ35
A
Appendix
Appendix 1 ✀ྡࢡࢭࢵࢩࣙࣥࢼࣥࣂ࣮
Gene name Accession number
Homo_sapiens_TET1 NM_030625 Pan_troglodytes_TET1 XM_507822 Gorilla_gorilla_TET1 XM_004049504 Pongo_abelii_TET1 XM_002820885 Nomascus_leucogenys_TET1 ENSNLEG00000011782 Macaca_fascicularis_TET1 XM_005565691 Papio_anubis_TET1 XM_003903859 Callithrix_jacchus_TET1 XM_002756304 Saimiri_boliviensis_TET1 XM_003928779 Ceratotherium_simum_TET1 XM_004427056 Canis_lupus_TET1 XM_536371 Felis_catus_TET1 ENSFCAG00000012673 Ailuropoda_melanoleuca_TET1 ENSAMEG00000018009 Pteropus_alecto_TET1 KB030800 Odobenus_rosmarus_TET1 XM_004393695 Orcinus_orca_TET1 XM_004280811 Trichechus_manatus_TET1 XM_004386395 Camelus_ferus_TET1 XM_006181184 Vicugna_pacos_TET1 XM_006212904 Capra_hircus_TET1 XM_005699119 Loxodonta_africana_TET1 XM_003409297 Ovis_aries_TET1 XM_004021627 Dasypus_novemcinctus_TET1 XM_004470524 Mustela_putorius_TET1 XM_004795559 Sus_scrofa_TET1 KC137683.1 Tupaia_chinensis_TET1 XM_006148936 Condylura_cristata_TET1 XM_004680833 Oryctolagus_cuniculus_TET1 XM_002718452 Otolemur_garnettii_TET1 XM_003783761 Spermophilus_tridecemlineatus_TET1 XM_005325968 Myotis_brandtii_TET1 XM_005876807
36 Heterocephalus_glaber_TET1 XM_004888511 Echinops_telfairi_TET1 XM_004701383 Jaculus_jaculus_TET1 XM_004657793 Cavia_porcellus_TET1 XM_003473584 Ictidomys_tridecemlineatus_TET1 ENSSTOG00000001527 Rattus_norvegicus_TET1 XM_006223879 Mus_musculus_TET1 NM_001253857 Mesocricetus_auratus_TET1 XM_005070904 Microtus_ochrogaster_TET1 XM_005360115 Panthera_tigris_TET1 XM_007095756 Balaenoptera_acutorostrata_TET1 XM_007167787 Leptonychotes_weddellii_TET1 XM_006728898 Physeter_catodon_TET1 XM_007121056 Bubalus_bubalis_TET1 XM_006070286 Chrysochloris_asiatica_TET1 XM_006835832 Myotis_davidii_TET1 XM_006760999 Homo_sapiens_TET2 NM_001127208 Pan_troglodytes_TET2 XM_003310400 Pan_paniscus_TET2 XM_003829967 Gorilla_gorilla_TET2 XM_004040224 Nomascus_leucogenys_TET2 ENSNLEG00000013289 Pongo_abelii_TET2 XM_002815025 Macaca_fascicularis_TET2 XM_005555594 Saimiri_boliviensis_TET2 XM_003929494 Callithrix_jacchus_TET2 XM_002745526 Mustela_putorius_TET2 XM_004748137 Canis_lupus_TET2 XM_535678 Orcinus_orca_TET2 XM_004269617 Loxodonta_africana_TET2 XM_003410366 Ailuropoda_melanoleuca_TET2 XM_002926670 Felis_catus_TET2 XM_006930937 Trichechus_manatus_TET2 XM_004380202 Odobenus_rosmarus_TET2 XM_004401875 Otolemur_garnettii_TET2 XM_003796334 Sus_scrofa_TET2 XM_003129278 Camelus_ferus_TET2 XM_006184616
37 Ceratotherium_simum_TET2 XM_004426613 Spermophilus_tridecemlineatus_TET2 XM_005339807 Tursiops_truncatus_TET2 XM_004319793 Oryctolagus_cuniculus_TET2 XM_002717196 Ovis_aries_TET2 XM_004009655 Myotis_davidii_TET2 XM_006778911 Chrysochloris_asiatica_TET2 XM_006874557 Jaculus_jaculus_TET2 XM_004662993 Echinops_telfairi_TET2 XM_004703292 Tupaia_chinensis_TET2 XM_006151157 Ochotona_princeps_TET2 XM_004594282 Chinchilla_lanigera_TET2 XM_005406228 Sarcophilus_harrisii_TET2 XM_003772926 Pteropus_alecto_TET2 KB031030 Monodelphis_domestica_TET2 XM_003341383 Microtus_ochrogaster_TET2 XM_005357312 Mesocricetus_auratus_TET2 XM_005081981 Bos_taurus_TET2 XM_001790146 Capra_hircus_TET2 XM_005681343 Octodon_degus_TET2 XM_004626707 Pantholops_hodgsonii_TET2 XM_005978869 Heterocephalus_glaber_TET2 XM_004866253 Rattus_norvegicus_TET2 XM_001077411 Mus_musculus_TET2 NM_001040400 Homo_sapiens_TET3 NM_001287491 Pongo_abelii_TET3 ENSPPYG00000012286 Macaca_fascicularis_TET3 XM_005575570 Canis_lupus_TET3 XM_005630554 Felis_catus_TET3 XM_003984133 Odobenus_rosmarus_TET3 XM_004398707 Equus_caballus_TET3 XM_001917114 Trichechus_manatus_TET3 XM_004369189 Chinchilla_lanigera_TET3 XM_005385530 Spermophilus_tridecemlineatus_TET3 XM_005329351 Lipotes_vexillifer_TET3 XM_007459392 Mustela_putorius_TET3 XM_004742217
38 Condylura_cristata_TET3 XM_004691540 Orcinus_orca_TET3 XM_004277087 Bubalus_bubalis_TET3 XM_006045954 Pantholops_hodgsonii_TET3 XM_005962644 Ceratotherium_simum_TET3 XM_004435517 Leptonychotes_weddellii_TET3 XM_006735303 Chrysochloris_asiatica_TET3 XM_006872404 Balaenoptera_acutorostrata_TET3 XM_007189011 Bos_mutus_TET3 XM_005897644 Sus_scrofa_TET3 XM_005662450 Heterocephalus_glaber_TET3 XM_004844710 Cavia_porcellus_TET3 XM_005003343 Octodon_degus_TET3 XM_004634101 Rattus_norvegicus_TET3 XM_006224966 Myotis_brandtii_TET3 XM_005869422 Microtus_ochrogaster_TET3 XM_005364657 Vicugna_pacos_TET3 XM_006218996 Mus_musculus_TET3 XM_006505773 Mesocricetus_auratus_TET3 XM_005071053 Myotis_davidii_TET3 XM_006760024 Peromyscus_maniculatus_TET3 XM_006994349 Jaculus_jaculus_TET3 XM_004668275 Echinops_telfairi_TET3 XM_004696098 Ochotona_princeps_TET3 XM_004590675 Sorex_araneus_TET3 XM_004619988 Elephantulus_edwardii_TET3 XM_006880851 Monodelphis_TET3 XM_007477889
39
A
45
A
51
