TABLE 1.
Translation of various mRNAs by S30 extracts derived from control and cold-shocked cells after different times of cold exposure
| mRNA | Translation temperature | S30 (control) | S30 Cs (30 min) | S30 Cs (60 min) | S30 Cs (90 min) | S30 Cs (120 min) |
| wtcspA | 37°C | 13.65 ± 0.60 | 27.60 ± 0.60 | 26.20 ± 0.70 | 19.70 ± 0.53 | 24.40 ± 1.70 |
| 15°C | 4.95 ± 0.28 | 7.56 ± 0.70 | 8.44 ± 0.38 | 14.19 ± 1.50 | 13.70 ± 0.60 | |
| cspA (Δ1-180) | 37°C | 15.03 ± 0.70 | 31.90 ± 0.40 | 25.90 ± 1.10 | 23.37 ± 1.10 | 22.60 ± 0.86 |
| 15°C | 3.35 ± 0.34 | 4.10 ± 0.66 | 3.58 ± 0.65 | 6.54 ± 0.65 | 7.51 ± 0.06 | |
| hns | 37°C | 2.07 ± 0.07 | 7.56 ± 0.23 | 6.58 ± 0.60 | 4.32 ± 0.23 | 5.30 ± 0.73 |
| 15°C | 0.82 ± 0.15 | 0.87 ± 0.19 | 0.76 ± 0.10 | 1.61 ± 0.16 | 1.62 ± 0.13 | |
| cspD | 37°C | 1.16 ± 0.06 | 2.27 ± 0.22 | n.d. | 2.70 ± 0.60 | 2.24 ± 0.40 |
| 15°C | 0.23 ± 0.08 | 0.00a | 0.00a | 0.00a | 0.00a | |
| hupA | 37°C | 2.29 ± 0.18 | 1.80 ± 0.25 | 1.44 ± 0.12 | 1.70 ± 0.05 | 1.76 ± 0.36 |
| 15°C | 0.29 ± 0.07 | 0.22 ± 0.05 | n.d. | 0.27 ± 0.06 | 0.58 ± 0.04 | |
| P4hupB | 37°C | 16.2 ± 0.50 | 26.30 ± 0.51 | 24.40 ± 0.51 | 19.19 ± 0.16 | 21.50 ± 1.30 |
| 15°C | 3.35 ± 0.42 | 3.82 ± 0.37 | 3.51 ± 0.32 | 5.73 ± 0.48 | 5.83 ± 0.42 | |
| P3hupB | 37°C | 12.80 ± 0.28 | 21.17 ± 0.36 | 19.50 ± 0.32 | 17.46 ± 0.24 | 19.40 ± 0.05 |
| 15°C | 1.85 ± 0.46 | 2.60 ± 0.13 | 2.30 ± 0.24 | 4.10 ± 0.29 | 4.10 ± 0.40 | |
| P2hupB | 37°C | 13.4 ± 0.28 | 17.89 ± 1.40 | 15.70 ± 0.52 | 16.32 ± 0.42 | 18.9 ± 0.15 |
| 15°C | 2.52 ± 0.34 | 2.46 ± 0.16 | 2.98 ± 0.40 | 3.95 ± 0.46 | 4.40 ± 0.50 |
The experiment was carried out as described in the text and Materials and Methods. As indicated in the headings, the reaction mixtures contained different types of cell extracts programmed with different mRNAs and were incubated at 37° or 15°C. Samples were taken at different time intervals until saturation was reached. The data were analyzed by the Regression Wizard program (SigmaPlot) and could be fit in all cases to an exponential equation (see text). The correlation coefficients R, which indicate how well the regression model describes the data, were found to be in all cases very close to the maximum theoretical value of 1, ranging from 0.9418 (hns mRNA translated at 37°C by control S30) to 0.9996 (cspA mRNA translated at 37°C by S30 60 min cs). The standard error of the estimate, which measures the actual variability about the regression plane of the underlying population, ranged from ±0.40 to ±1.70.
The results presented in the table refer to the number of pmoles of product synthesized after 15 min incubation (± the standard error).
n.d. = not done.
aBarely detectable above background level.