Table 1 |
||||
|
Alanine scan of LEXA-HSF1 |
||||
|
Construct |
Relative luciferase |
|||
|
|
||||
|
C |
SD |
HS |
SD |
|
|
|
||||
|
LEXA-HSF1 |
0.50 |
0.1 |
100 |
13.5 |
|
LEXA-HSF1(F) |
0.32 |
0.1 |
100 |
6.3 |
|
T97A |
0.83 |
0.3 |
122 |
5.0 |
|
T120A/S121A(F) |
0.59 |
0.1 |
115 |
4.7 |
|
S123A/T124A |
0.65 |
0.6 |
95 |
6.6 |
|
S127A |
0.50 |
0.0 |
84 |
5.1 |
|
S136A/T138A(F) |
0.23 |
0.1 |
112 |
11.6 |
|
T142A(F) |
0.22 |
0.0 |
83 |
11.0 |
|
S156A(F) |
0.29 |
0.0 |
102 |
6.1 |
|
S(174/195/199)A |
0.30 |
0.0 |
105 |
8.2 |
|
S216A |
0.52 |
0.1 |
77 |
10.5 |
|
S218A(F) |
0.31 |
0.0 |
117 |
9.5 |
|
S221A(F) |
0.64 |
0.4 |
70 |
8.3 |
|
Y225A/S(226/230)A(F) |
0.39 |
0.0 |
92 |
5.8 |
|
S(237/241/244)A/Y240A |
0.54 |
0.1 |
126 |
6.4 |
|
Y247A/S(248/249)A(F) |
0.72 |
0.1 |
127 |
2.2 |
|
S(250/251)A/Y253(F) |
1.21 |
0.3 |
99 |
16.6 |
|
S(260/261)A |
0.84 |
0.2 |
119 |
19.1 |
|
S266A/T269A(F) |
0.95 |
0.3 |
83 |
11.8 |
|
S275A |
1.05 |
0.3 |
75 |
5.5 |
|
S279A |
0.51 |
0.0 |
103 |
24.7 |
|
S290A |
0.32 |
0.0 |
101 |
13.5 |
|
S291A |
1.43 |
0.4 |
80 |
4.1 |
|
S292A |
1.47 |
0.6 |
84 |
3.1 |
|
S303A |
0.97 |
0.3 |
81 |
13.9 |
|
S307A(F) |
0.36 |
0.0 |
148 |
33.7 |
|
S(303/307)A(F) |
1.27 |
0.5 |
249 |
16.9 |
|
S314A(F) |
0.34 |
0.0 |
131 |
18.3 |
|
S(319/320)A/T323A |
0.83 |
0.2 |
78 |
10.8 |
|
S326A/T328A |
2.15 |
1.0 |
46 |
1.9 |
|
S333A(F) |
0.37 |
0.0 |
103 |
9.3 |
|
S338A(F) |
0.51 |
0.1 |
88 |
10.1 |
|
S344A(F) |
0.47 |
0.1 |
101 |
13.4 |
|
T346A(F) |
0.60 |
0.1 |
108 |
15.0 |
|
T349A(F) |
0.35 |
0.1 |
95 |
12.6 |
|
T(355/357)A(F) |
0.34 |
0.0 |
129 |
12.6 |
|
S(363/368)A/T(367/369)A(F) |
1.05 |
0.6 |
92 |
14.4 |
|
S375A |
2.13 |
1.9 |
73 |
13.6 |
|
S385A(F) |
0.57 |
0.1 |
121 |
5.7 |
|
S393A |
1.38 |
0.6 |
83 |
14.9 |
|
T400A/S(403/404)A(F) |
0.57 |
0.2 |
63 |
2.9 |
|
T411A/S412A(F) |
0.57 |
0.0 |
85 |
6.3 |
|
S(419/421)A/T423A(F) |
0.48 |
0.0 |
101 |
6.7 |
|
S428A |
0.88 |
0.1 |
75 |
9.6 |
|
S(434/435/438)A |
1.39 |
0.1 |
81 |
12.9 |
|
S444A |
1.27 |
0.2 |
88 |
13.2 |
|
S(457/458/461)A(F) |
0.49 |
0.1 |
7 |
0.3 |
|
Y468A/T469A |
2.12 |
1.9 |
103 |
16.6 |
|
S(480/485)A/T483A |
0.47 |
0.1 |
80 |
4.0 |
|
S(498/501)A/Y499A |
1.08 |
0.1 |
80 |
9.2 |
|
T511A/S513A(F) |
0.44 |
0.1 |
22 |
1.0 |
|
T516A/S518A(F) |
0.87 |
0.1 |
78 |
8.1 |
|
T527A/S529A |
1.14 |
0.2 |
91 |
4.6 |
|
S121A |
1.73 |
0.2 |
106 |
12.1 |
|
S230A |
1.64 |
0.3 |
101 |
22.7 |
|
S319A |
0.60 |
0.1 |
105 |
14.3 |
|
S320A |
0.36 |
0.1 |
91 |
12.8 |
|
T323A |
1.97 |
0.5 |
102 |
17.6 |
|
S326A |
1.66 |
0.3 |
40 |
8.1 |
|
S419A |
1.67 |
0.2 |
104 |
17.8 |
|
|
||||
|
C: not-heated; SD: standard deviation; HS: heat-treated. Transactivation assays were carried out as discussed in Methods and Results. (F) refers to the presence of an amino-terminal FLAG tag (see Methods). The activities of FLAG-tagged mutants are expressed relative to the activity of FLAG-LEXA-HSF1, and those of not-FLAG-tagged mutants to the activity of LEXA-HSF1. |
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|
Guettouche et al. BMC Biochemistry 2005 6:4 doi:10.1186/1471-2091-6-4 |
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