Article Figures & Data
Figures
- Fig. 1.
In vivo transcription from the alkA promoter. The effects of the substitutions in ς70 region 4 onalkA expression are shown as percentages of alkAexpression with wild-type ς70. Values (± standard deviations) are averages of four independent experiments. The average value for the wild type was 342 Miller units.
- Fig. 2.
In vitro transcription with reconstituted RNA polymerases (50 nM). Lanes 1 to 3, wild-type Eς70; lanes 4 to 6, Eς70KA593; lanes 7 to 9, Eς70KA597; lanes 10 to 12, Eς70RA603. Lanes 1, 4, 7, and 10, no Ada protein; lanes 2, 5, 8, and 11, 0.2 μM unmethylated Ada; lanes 3, 6, 9, and 12, 0.2 μM methylated Ada. The positions of the mainlacUV5 and alkA transcripts are indicated.
- Fig. 3.
Ratio of alkA/lacUV5 transcripts. Values are percentages of transcription by wild-type Eς70 in the presence of meAda and are averages of three independent experiments. Dark grey bars, transcription levels in the absence of Ada; light grey bars, transcription levels in the presence of unmethylated Ada; black bars, transcription levels in the presence of methylated Ada. Standard deviations were less than 15%.
- Fig. 4.
Gel retardation assays performed with reconstituted RNA polymerase. Odd-numbered lanes, no meAda; even-numbered lanes, 0.2 μM meAda. Lanes 1 and 2, no RNA polymerase; lanes 3 and 4, wild-type Eς70; lanes 5 and 6, Eς70KA593; lanes 7 and 8, Eς70KA597; lanes 9 and 10, Eς70RA603. F, unbound alkApromoter DNA; C, RNA polymerase-alkA complex.
- Fig. 5.
In vitro transcription with Eς70 (lanes 1 to 3) and EςS (lanes 4 to 6) forms of RNA polymerase. Lanes 1 and 4, no Ada protein; lanes 2 and 5, 0.2 μM Ada; lanes 3 and 6, 0.2 μM meAda. The positions of the mainlacUV5 and alkA transcripts are indicated.
- Fig. 6.
Gel retardation assays performed with Eς70and EςS forms of RNA polymerase. Odd-numbered lanes, nomeAda; even-numbered lanes, 0.2 μM meAda. Lanes 1 and 2, no RNA polymerase; lanes 3 and 4, Eς70; lanes 5 and 6, EςS. F, unbound alkA promoter DNA; C70 and CS, Eς70-alkA and EςS-alkA complexes, respectively.
