Tanaka A, Hirano H, Kikuchi M, Kitayama S, Watanabe H
Biotechnology Laboratory, Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, Gunma, Japan.
Radiat Environ Biophys. 1996 May;35(2):95-9. doi: 10.1007/BF02434031.
In order to examine radiation-induced proteins in an extremely radioresistant bacterium, Deinococcus radiodurans R1, changes in cellular proteins after gamma-irradiation were analysed by two-dimensional gel electrophoresis and silver staining. Nine proteins (190, 120, 87,60, 58, 52, 46, 41 and 41 kDa) were increased (or appeared) and more than 13 proteins diminished after gamma-irradiation at 6 kGy. Increase of eight proteins (except for 190-kDa protein) was prevented when the cells were irradiated in the presence of chloramphenicol. Three proteins, 87, 60 and 46 kDa, continued to be synthesized during post-irradiation incubation, and the amounts of these proteins increased with higher doses in a range of 1-12 kGy. Changes in the amount of proteins after irradiation in the R1 strain were compared with those in a moderately radioresistant mutant (rec I) and in a highly radiosensitive mutant (rec30). These three proteins were increased in both R1 and recI, but not in rec 30, suggesting that they are characteristic for radioresistant strains. In addition, from the microsequence analysis, the 46-kDa protein was found to be homologous to the EF-Tu protein of Escherichia coli, whereas the remarkable homologous sequence to the N-terminal of the 60-kDa protein was not found among the known proteins.
为了检测极端耐辐射细菌耐辐射球菌R1中辐射诱导的蛋白质，通过二维凝胶电泳和银染分析了γ射线照射后细胞蛋白质的变化。9种蛋白质（190、120、87、60、58、52、46、41和41 kDa）在γ射线6 kGy照射后增加（或出现），13种以上蛋白质减少。当细胞在氯霉素存在下照射时，8种蛋白质（除190 kDa蛋白质外）的增加受到抑制。3种蛋白质，87、60和46 kDa，在照射后孵育期间持续合成，并且在1-12 kGy范围内随着剂量增加这些蛋白质的量也增加。将R1菌株照射后蛋白质含量的变化与中度耐辐射突变体（rec I）和高度辐射敏感突变体（rec30）中的变化进行比较。这3种蛋白质在R1和recI中均增加，但在rec 30中未增加，表明它们是耐辐射菌株的特征。此外，通过微序列分析，发现46 kDa蛋白质与大肠杆菌的EF-Tu蛋白质同源，而在已知蛋白质中未发现与60 kDa蛋白质N端显著同源的序列。
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