Honda Takashi, Hara Tadashi, Nan Jinghua, Zhang Xjaodong, Kimura Makoto
Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University, Fukuoka, Japan.
Biosci Biotechnol Biochem. 2010;74(2):266-73. doi: 10.1271/bbb.90550. Epub 2010 Feb 7.
We examined the functional equivalency between Escherichia coli RNase P protein (C5) and Pyrococcus horikoshii RNase P proteins (PhoPop5, PhoRpp21, PhoRpp29, PhoRpp30, and PhoRpp38) for RNase P activity. The C5 protein and P. horikoshii RNase P proteins were unable to activate non-congnate RNase P RNAs, P. horikoshii RNase P RNA (PhopRNA) and E. coli RNase P RNA (M1 RNA) respectively. Two chimeric RNAs, in which functional C- and S-domains of M1 RNA and PhopRNA were exchanged, were prepared and characterized with respect to the cleavage of P. horikoshii pre-tRNA(Tyr) in the presence of C5 or P. horikoshii proteins. The results suggest that PhoPop5 and PhoRpp30 function equivalently to the C5 protein in the E. coli RNase P, being involved in activation of the PhopRNA C-domain. On the other hand, PhoRpp21 and PhoRpp29 are implicated in stabilization of the PhopRNA S-domain.
我们检测了大肠杆菌核糖核酸酶P蛋白（C5）与嗜热栖热菌核糖核酸酶P蛋白（PhoPop5、PhoRpp21、PhoRpp29、PhoRpp30和PhoRpp38）在核糖核酸酶P活性方面的功能等效性。C5蛋白和嗜热栖热菌核糖核酸酶P蛋白分别无法激活非同源核糖核酸酶P核糖核酸，即嗜热栖热菌核糖核酸酶P核糖核酸（PhopRNA）和大肠杆菌核糖核酸酶P核糖核酸（M1 RNA）。制备了两种嵌合核糖核酸，其中M1 RNA和PhopRNA的功能性C结构域和S结构域进行了交换，并在C5或嗜热栖热菌蛋白存在的情况下，针对嗜热栖热菌前体tRNA（Tyr）的切割进行了表征。结果表明，PhoPop5和PhoRpp30在大肠杆菌核糖核酸酶P中的功能与C5蛋白等效，参与PhopRNA C结构域的激活。另一方面，PhoRpp21和PhoRpp29与PhopRNA S结构域的稳定有关。
