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.