6Ü D ÄeIF4E RNA High affinity RNA for mammalian...
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ㄵහᐖࡡさ蹇 ハ貅嵃┘困9ゼ麻慚嗔攷 eIF4E 作0倫載三 RNA 仕字獅治質作鮫三9ゼ揆朖㌣└ 搾よ躬埼材搾愈滿斟ɔ眦ブ笹搾槐麻坤 High affinity RNA for mammalian translation initiation factor 4E ; A novel inhibitor of translation. 翔琳ǖ 鐇 鏗 ㌞ 繈 ǚ煖Ą ヂH搾梅挺攷oI作昨┞惚鷺呆癪㌣└栽敍圜載三腰単崎薩9ゼ里眦搾呆癪昨 o73& 搾㌹父里眦埼燐┞漉 梅挺攷搾oI蕕作├藜疽堠載三財殺漉る杤鷺呆癪㌣└搾薩埼作耆晒イ坂漉禁ヤ搾嵜挽疾返杢作臍さ鷺痙 房桟艱堺咲最三腰 ┷饋ヂH搾9ゼ呆癪崎昨漉9ゼ麻慚嗔攷 g.+6* 作鮫三 o73& 搾 7)鑠❻搾o 9 ,rrr3 ∑姉示 紙字└『∭搾をか埼漉焉榊採搾g.+ 搾超圜作鮫堺咲鹿次死質璽 628 指児自試紙視栽 o73& 作イ椁斎傘三謹 8栽臍さ崎催三∑嘸θ∭腰g.+6* 埼姉示紙字搾0倫昨砦搾謹8搾十滝鑓网崎催晒漉鹿叱囁杢皐 6*/'kpIkpi 5tqvgkpu 作鮫三乱杤鷺呆癪桟略犀咲最三腰撮財 g.+6* 昨捗搾 g.+ 搾単崎薩禁ヤ敏崎材搾返攷豁栽鎬薩椈 鷺最嗔攷崎催晒漉砦搾oI蕕搾嵜⎫栽禁ヤ搾o絆作彖歳鷺疽堠桟濁宰三砦埼栽å皿傘咲最三腰凍宰朔漉 g.+6* 桟謹忘oI菜財禁ヤ昨倁由ゼɔ鷺返◎桟災晒ǒ菜4⑾杢載三腰ヂ鉄敏崎昨 g.+6* 搾oI蕕嵜本栽 歯視馳⒬漉ソ⒬漉彖4⒬禁ヤ単作巠リ作擦皿傘咲最三腰撮財漉g.+6* 搾oI蕕搾嵜本作程堺咲捗搾浴⒬ 梅挺攷ドH栽謹忘oI載三腰砦搾鮫塞作禁ヤ搾⒬杢埼 g.+6* 搾oI蕕搾嵜本栽杤藜作門惨堺咲最三砦埼 栽1倔斎傘咲歳咲最三腰 )3&漉 73& 埼最堺財饋囁昨ヂH搾梅挺攷筅寃埼挺祉挂鉄埼菜咲å皿傘咲最三栽漉返攷脈┞柱貼ブ作才最 咲薩玫最Qれ磔疾㌣勝磔桟薩榊返攷崎催三腰ǐ烱漉砦傘皿搾ヂ鉄返攷桟某や桟昨裁殺埼菜財x煦眦ブ笹 埼槐麻載三贐菜最モ擦栽鷺斎傘咲最三腰.p [kvtq ugngHvkqp 叛作鮫晒突舍斎傘財漉催三╇ɔH㈻作梏菜 咲Qれɔ作0倫載三返攷桟仕字獅治質埼最塞腰仕字獅治質昨弩災搾寉倫漉╇ɔH㈻搾瓷堀鷺仕叱獅思試 施視埼菜咲㌣勝載三財殺漉┎蔗作門〛菜財獅叱飼子㈻桟獅質師紙視埼載三砦埼作鮫晒漉仕字獅治質搾愈 や仂笹搾眦ブ栽鐵瘋斎傘三腰鑢ŠW崎昨漉⒬禁ヤ作擦皿傘三 g.+6* 搾謹忘oI作鮫三禁ヤ搾⒬杢搾臟呆 桟┘ɔ埼菜咲漉g.+6* 作Qれɔ作0倫載三仕字獅治質桟突舍菜漉材搾㌣勝磔搾よ隸桟〒堺財腰
Transcript of 6Ü D ÄeIF4E RNA High affinity RNA for mammalian...
![Page 1: 6Ü D ÄeIF4E RNA High affinity RNA for mammalian ...gakui.dl.itc.u-tokyo.ac.jp/data/h17/121070/121070a.pdf · 15 ] Ä aさ 15 ]8 ┘ *sゼ6Ü D ÄeIF4E ^) L ~RNA È ° Ï í ^ {](https://reader031.fdocument.pub/reader031/viewer/2022012008/611e83e228afe31f385a52b1/html5/thumbnails/1.jpg)
さ
┘ ゼ eIF4E RNA ゼ よ
High affinity RNA for mammalian translation initiation factor 4E ; A novel inhibitor of translation.
ゼ 73& ㌹
├ さ
┷ ゼ ゼ .+ * 73& ❻ , 3
.+ 8 73&
さ .+ * * ' I
5 .+ * .+
.+ * ⑾ .+ *
⒬ ⒬ ⒬ .+ * ⒬
⒬ .+ *
)3& 73& ┞
モ . [ H ㈻
㈻
㈻
⒬ .+ * ⒬
┘ .+ * よ
![Page 2: 6Ü D ÄeIF4E RNA High affinity RNA for mammalian ...gakui.dl.itc.u-tokyo.ac.jp/data/h17/121070/121070a.pdf · 15 ] Ä aさ 15 ]8 ┘ *sゼ6Ü D ÄeIF4E ^) L ~RNA È ° Ï í ^ {](https://reader031.fdocument.pub/reader031/viewer/2022012008/611e83e228afe31f385a52b1/html5/thumbnails/2.jpg)
73& 9 F I , I 8H H * F I 8_ F 3F
[ H 73&
- .+ * 73& 3 39& I \
㌹ 5(7 73&
857 よ .+ *
┞ よ モ F F .+ *
0I " 2 .+ * .+ *
HF F F ,95 F
I \ F F .+ * HF
F F .+ *
ゼ
[ ゼ ゼ ッ (&9 1:(
-(; .7*8 - F ( ; . F 7 G
* ^ 8 ❻ HF 73& ゼ -(; .7*8
.+ 8 ├ 47+ ゼ
73& (&9 1:( ゼ
73& 8 2 [ ゼ F F
2 1:( ゼ (&9 ゼ
F F 3 F I
(&9 1:( ゼ F F .+ *
さ ゼ
& F 73& F F
,
F F
.+ * F F
.+ * F F 857 よ
100 114 102 103 91 54 53 21 15 CAT/LUC (%)
CAT LUC
Cap-dependent Cap-independent
HCV IRES Cap (eIF4E-independent) (eIF4E-dependent)
No RNA 0.5 1.0 2.0 5.0 0.5 1.0 2.0 5.0 (µM)
N40 random Aptamer 1
CAT
LUC
[ ゼ
ゼ .+ *
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よ 7 & F F
0 &
F F .+ * ┞ & 1^ さ
F F
.+ *
.+ *
ッ ッ
さ .+ *
┷ 8
H [ F .+ * .+ *
.+ * F F モ
.+ * .+ *
.+ * (>( .+ * '>
(IH - (>( * ^ *
F F 73&
73&
F FI G _^ 73& ス
スゝ 73&
さ 7_&
7_& " 73& スゝ
7_& ,&1 (>( 78
.+ * .+ *
'> HIH -.8 (>( .+ * ^ .+ * ㈻㌹
.+ * 7_& 73& 7_&
Lys206
Arg112
-50
0
50
100
150
-20 0 20 40 60 80 100 120 140
R112A RUs
Time (sec) -50
0
50
100
150
-20 0 20 40 60 80 100 120 140
K206A RUs
Time (sec) -50
0
50
100
150
-20 0 20 40 60 80 100 120 140
Wild RUs
Time (sec)
eIF4E (0.05 µM)
eIF4E (0.1 µM)
eIF4E (0.15 µM)
eIF4E (0.2 µM)
eIF4E (0.5 µM)
857よ .+ *
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.+ * 73& .+ * F F
F F .+ * ┞ & 1^ .+ *
73&
┞ ┞ [ H さ さ
= よ 9 F / 2 ' , & ┘
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┘ ┞ 9 1^ 8
┞ 73&
73& 73&
& F ゼ .+ *
F F 7_& .+ *
ふ F F .+ *
ゼ 7_& 73& ㈻
.+ * さ 73& F F ㈻
73& ぞ 73&
Ⅴ 73&
pRS416
Pro Human eIF4E
ter
ARS4/CEN6
HIS3 marker
URA3
AmpR Yeast eIF4E
heIF4E yeIF4E
pRS416
GAL pro (RzAp1)n
2µ ori
TRP1
AmpR
ter
Ribozyme Ribozyme
Aptamer 1
cleavage
n (n=1, 2, 4, 8)
.+ *
8( , H Υ 8( ,F Υ
Vector
RzAp1
(RzAp1)2
(RzAp1)4
(RzAp1)8
Vector
RzAp1
(RzAp1)2
(RzAp1)4
(RzAp1)8
[Yeast eIF4E] [Human eIF4E]
Aptamer 1
