Revealing the Mechanism of HSP104 Transcription Initiation in the ...

Table 4. Strains required for positive regulation of HSP104.
β-Gal activity (% of WT) β-Gal activity (% of WT) β-Gal activity (% of WT)
Strain Cofatctor complex Uninduced Induced Uninduced Induced Uninduced Induced
HSP104 HSP104 STRE-LacZ STRE-LacZ HSE-LacZ HSE-LacZ
bdf1∆ TFIID 225 135 742 122 46 67
elp3∆ HAT 116 185 227 140 156 111
chd1∆ Homodimer 74 156 143 152 149 84
caf1∆ CCR4-NOT 227 60 237 87 15 13
dhh1∆ CCR4-NOT 319 67 873 132 17 7
srb9∆
SRB/MED,CCR4-
NOT
150 59 723 104 52 38
med1∆ SRB/MED 154 107 715 229 42 38
med9∆ SRB/MED 179 93 1550 484 29 32
nut1∆ SRB/MED 135 68 580 249 101 78
β-Gal activity (% of WT) β-Gal activity (% of WT) β-Gal activity (% of WT)
Strain Cofatctor complex Uninduced Induced Uninduced Induced Uninduced Induced
HSP104 HSP104 STRE-LacZ STRE-LacZ HSE-LacZ HSE-LacZ
gcn5∆ ADA,SAGA 23 69 99 17 13 20
spt7∆ SAGA 58 16 325 37 6 8
sas3∆ NuA3 3 7 0 2 7 6
hos2∆ HDAC 36 64 33 55 35 26
sir2∆ HDAC 38 71 29 77 38 46
hst1∆ HDAC 36 55 80 105 72 56
snf6∆ SWI/SNF 13 50 0 0 10 23
srb10∆
SRB/MED,CCR4-
NOT
72 15 469 41 13 6
srb2∆ SRB/MED 33 110 154 69 32 94
srb5∆ SRB/MED 10 23 78 23 12 29
srb8∆ SRB/MED 60 28 614 90 69 28
dst1∆ TFIIS 25 36 182 111 58 40
rpb4∆ RNA PolII core 2 0.4 17 0.6 1 0.4
cdc73∆ Paf1 complex 11 20 11 17 4 12
leo1∆ Paf1 complex 21 29 37 51 34 28
hpr1∆ Paf1,THO/TREX 0.1 1 0 1.5 0 0.1
mft1∆ THO/TREX 4 3 5 0.1 0.4 0.5
Table 5. Strains which induced an up-regulation of HSP104 (i.e. downregulate
HSP104).
In order to confirm that the decreased or increased reporter activities measured
in the mutants reflect indeed defects in expression of endogenous HSP104, we
performed S1 analysis to detect endogenous mRNA levels of HSP104. We first
measured RNA levels in strains which demonstrated a decrease in the β-galactosidase
activity of -334LacZ. Most strains listed in Table 4 indeed had lower RNA levels
40