6Ü d Äeif4e rna high affinity rna for mammalian...
TRANSCRIPT
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
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 .+ *
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 .+ *
.+ * 73& .+ * F F
F F .+ * ┞ & 1^ .+ *
73&
┞ ┞ [ H さ さ
= よ 9 F / 2 ' , & ┘
& .+ * 9 1^ 8 ┞ 1^
┘ ┞ 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