Table 2.
DNA binding competition assay with mutant FeaR binding sites
| Competitora | DNA sequenceb | IC50 (nM)c |
|---|---|---|
| Site 1 | ATGAAAAGCTGGCACACCTGCCAAACCCCCT | 660 (50) |
| Site 1-A1G | ATGAAgAGCTGGCACACCTGCCAAACCCCCT | 910 (51) |
| Site 1-A1C | ATGAAcAGCTGGCACACCTGCCAAACCCCCT | 1,070 (46) |
| Site 1-A2C | ATGAAAcGCTGGCACACCTGCCAAACCCCCT | 1,330 (65) |
| Site 1-C4G | ATGAAAAGgTGGCACACCTGCCAAACCCCCT | 700 (40) |
| Site 1-T5G | ATGAAAAGCgGGCACACCTGCCAAACCCCCT | No competition |
| Site 1-G6T | ATGAAAAGCTtGCACACCTGCCAAACCCCCT | No competition |
| Site 1-C8A | ATGAAAAGCTGGaACACCTGCCAAACCCCCT | No competition |
| Site 1-A9C | ATGAAAAGCTGGCcCACCTGCCAAACCCCCT | No competition |
| Site 1-A11C | ATGAAAAGCTGGCACcCCTGCCAAACCCCCT | 1,570 (106) |
| Site 1-C13A | ATGAAAAGCTGGCACACaTGCCAAACCCCCT | 691 (47) |
| Site 1-T14G | ATGAAAAGCTGGCACACCgGCCAAACCCCCT | 2,000 (82) |
| Site 1-G15T | ATGAAAAGCTGGCACACCTtCCAAACCCCCT | 1,810 (85) |
| Site 1-C17T | ATGAAAAGCTGGCACACCTGCtAAACCCCCT | 1,120 (60) |
| Site 1-A18T | ATGAAAAGCTGGCACACCTGCCtAACCCCCT | 4,120 (362) |
| Site 1-A19T | ATGAAAAGCTGGCACACCTGCCAtACCCCCT | 1,370 (86) |
| Site 1-A20C | ATGAAAAGCTGGCACACCTGCCAAcCCCCCT | 5,800 (517) |
| Site 2 | ATGAAAACCTGGCAGGTGCAGGCAATCCCCT | 580 (43) |
| Site 2-A23C | ATGAAAcCCTGGCAGGTGCAGGCAATCCCCT | 950 (32) |
| Site 2-C25G | ATGAAAACgTGGCAGGTGCAGGCAATCCCCT | 650 (54) |
| Site 2-T26G | ATGAAAACCgGGCAGGTGCAGGCAATCCCCT | No competition |
| Site 2-G27T | ATGAAAACCTtGCAGGTGCAGGCAATCCCCT | No competition |
| Site 2-C29A | ATGAAAACCTGGaAGGTGCAGGCAATCCCCT | No competition |
| Site 2-A30C | ATGAAAACCTGGCcGGTGCAGGCAATCCCCT | 5,010 (361) |
| Site 2-G32C | ATGAAAACCTGGCAGcTGCAGGCAATCCCCT | 2,650 (156) |
| Site 2-A36C | ATGAAAACCTGGCAGGTGCcGGCAATCCCCT | 1,840 (101) |
| Site 2-A40C | ATGAAAACCTGGCAGGTGCAGGCcATCCCCT | 7,846 (700) |
| Site 2-A41C | ATGAAAACCTGGCAGGTGCAGGCAcTCCCCT | No competition |
a
Promoter-proximal (site 1) and -distal (site 2) FeaR binding sites from the tynA promoter. Nucleotides are numbered according to the sequence logo (Fig. 3A).
b
The site 1 and site 2 sequences are underlined, and mutations are in lowercase.
c
Numbers in parentheses are errors estimated from the data fitting.