Table S2. Plasmids used in this study.
Plasmid
pM128
(1-1-1-1)
pM114
(1-1-4-4)
pVM35
(4-4-4-4)
pVM36
(4-4-1-1)
pVM15
(DEL6)
pVM14
(DEL16)
pVM13
(DEL26)
pVM12
(DEL36)
pVM11
(DEL46)
pVM16
(DELup39)
Description
low copy URA3 vector containing uORF1 only at its original position (350 nt
from GCN4-lacZ), with HindIII restriction site upstream of uORF1
low copy URA3 vector containing uORF1 only (with HindIII upstream); the
coding sequence and 25 nt of its 3' flanking sequence was replaced by the
corresponding sequences of uORF4
low copy URA3 vector containing uORF1 only (with HindIII upstream); uORF1’s
5’ flanking sequences (nt -181 to -1), the coding region, and the 3’ flanking
sequences (25 nt beyond the stop codon) were replaced by the corresponding
sequences of uORF4
low copy URA3 vector containing uORF1 only only (with HindIII upstream); );
uORF1’s 5’ flanking sequences (nt -181 to -1) were replaced by the
corresponding sequences of uORF4
low copy URA3 vector containing uORF1 only at its original position; the
sequences -21 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -31 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -41 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -51 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -61 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -181 to -143 upstream of uORF1 were deleted
Source of
reference
[4]
[5]
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[2]
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pVM18
(DEL56)
pVM20
(DEL76)
pVM21
(DEL109)
pVM45
(SUB31)
pVM46
(SUB41)
pVM47
(SUB49)
pVM26
(DELII)
pVM27
(CAAII)
pVM31
(AA-C)
pVM50
(DEL36+AA-C)
pVM80
(G4-uORF1)
pVM91
(Y1-uORF1)
low copy URA3 vector containing uORF1 only at its original position; the
sequences -71 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -91 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -125 to -16 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequence -40 to -32 upstream of uORF1 were substituted by complementary
sequences
low copy URA3 vector containing uORF1 only at its original position; the
sequences -49 to -41 upstream of uORF1 were substituted by complementary
sequences
low copy URA3 vector containing uORF1 only at its original position; the
sequences -54 to -50 upstream of uORF1 were substituted by complementary
sequences
low copy URA3 vector containing uORF1 only at its original position; the
sequences -76 to -55 upstream of uORF1 were deleted
low copy URA3 vector containing uORF1 only at its original position; the
sequences -76 to -55 upstream of uORF1 were substituted by a stretch of CAA
triplets of the identical length to the original sequence
low copy URA3 vector containing uORF1 only at its original position; G-129A,
C-128A and G-109C substitutions were inserted upstream of uORF1
low copy URA3 vector containing uORF1 only at its original position; the
uORF1’s mutations DEL36 and AA-C were combined
low copy URA3 vector containing uORF1 only at its original position (350 nt
from GCN4-lacZ) with SacII restriction site upstream of uORF1
low copy URA3 vector containing uORF1 only at its original position; the 5’ UTR
of uORF1 of GCN4 (-229_-9) was replaced by the corresponding sequence of
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YAP1 (-81_-9)
pVM92
low copy URA3 vector containing uORF1 only at its original position; the 5’ UTR
of uORF1 of GCN4 (-229_-9) was replaced by the corresponding sequence of
(Y2-uORF1)
YAP2 (-101_-9)
low copy URA3 vector containing the YAP1 gene with its complete 5’ UTR
pVM95
fused with lacZ under control of the GCN4 promoter
(Y1-lacZ)
low copy URA3 vector containing the YAP2 gene with its complete 5’ UTR
pVM96
fused with lacZ under control of the GCN4 promoter
(Y2-lacZ)
pVM97
a derivative of pVM91; C-32G and G-33G substitutions were inserted upstream
(Y1-uORF1-“stem”_C-32G G-33C) of uORF1
pVM98
derivative of pVM91, G-45U and C-57A substitutions were inserted upstream of
(Y1-uORF1-hairpin_G-45U C-57A) uORF1
low copy URA3 vector containing wild-type GCN4 leader
p180
(YCp50–GCN4–lacZ)
low copy URA3 vector containing uORF1 only placed 32 nt from GCN4-lacZ
pG67
pM199
low copy URA3 vector containing uORF1 only at the position of uORF4 (140 nt
from GCN4-lacZ)
p209
low copy URA3 vector containing uORF1 only at its original position (350 nt
from GCN4-lacZ)
pM23 (supp info)
low copy URA3 vector containing uORF1 and uORF4 only at their original
#
positions relative to GCN4-lacZ
(wt ; 1-1-1-1)
pVM55 (supp info)
derivative of pM23; mutation DEL16 introduced upstream of uORF1
(DEL16)
pVM54 (supp info)
derivative of pM23; mutation DEL36 introduced upstream of uORF1
(DEL36)
pVM59 (supp info)
derivative of pM23; mutation SUB49 introduced upstream of uORF1
(SUB49)
pVM60 (supp info)
derivative of pM23; deletion of a double-circle hairpin (nt -129 to -83) upstream
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[10]
[11]
[11]
[4]
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(DELhairpin)
pVM61(supp info)
(DELII)
pVM56 (supp info)
(CAAII)
pVM53 (supp info)
(DEL46)
pVM52 (supp info)
(DELup39)
pVM37 (supp info)
(bg#; 4-4-1-1)
YCplac111
of uORF1
derivative of pM23; mutation DELII introduced upstream of uORF1
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derivative of pM23; mutation CAAII introduced upstream of uORF1
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derivative of pM23; mutation DEL46 introduced upstream of uORF1
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derivative of pM23; mutation DELup39 introduced upstream of uORF1
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derivative of pM23; mutation 4-4-1-1 introduced upstream of uORF1
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single-copy cloning vector, LEU2
[7]
pRS-a/TIF32-His
low-copy a/TIF32-His in LEU2 plasmid, from pRS315
[6]
YCp-a/TIF32-His-L
YCp-a/TIF32-His-screen
single-copy a/TIF32-His in LEU2 plasmid, from YCplac111
single-copy a/TIF32-His with BamHI and NdeI sites introduced just in front of
the start codon of a/TIF32 in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box6-His in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box8-His in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box17- His in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box6+8- His in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box6+17- His in LEU2 plasmid, from YCplac111
single-copy a/tif32-Box8+17- His in LEU2 plasmid, from YCplac111
cloning vector for GAL4 activation domain fusion followed by T7 promoter,LEU2
a/TIF32-NTD cloned under T7 promoter, from pGADT7
a/tif32-NTD-Box6 cloned under T7 promoter, from pGADT7
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YCp-a/tif32-Box6-His
YCp-a/tif32-Box8-His
YCp-a/tif32-Box17-His
YCp-a/tif32-Box6+8-His
YCp-a/tif32-Box6+17-His
YCp-a/tif32-Box8+17-His
pGADT7 (supp info)
pGAD-a/TIF32-NTD (supp info)
pGAD-a/tif32-NTD-Box6 (supp
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CLONTECH
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info)
pGAD- a/tif32-NTD-Box17 (supp
info)
pGAD- a/tif32-NTD-Box6+17
(supp info)
pGEX-RPS0A (supp info)
a/tif32-NTD-Box17 cloned under T7 promoter, from pGADT7
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a/tif32-NTD-Box6+17 cloned under T7 promoter, from pGADT7
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GST-RPS0A fusion plasmid from pGEX-4T-1
I.D. and L.V.
unpublished
data