Ameres Stefan L, Shcherbakov Dmitry, Nikonova Ekaterina, Piendl Wolfgang, Schroeder Renée, Semrad Katharina
Max F. Perutz Laboratories, Department of Biochemistry, University of Vienna, Dr Bohrgasse 9/5, A-1030 Vienna, Austria.
Nucleic Acids Res. 2007;35(11):3752-63. doi: 10.1093/nar/gkm318. Epub 2007 May 21.
RNA chaperone activity is defined as the ability of proteins to either prevent RNA from misfolding or to open up misfolded RNA conformations. One-third of all large ribosomal subunit proteins from E. coli display this activity, with L1 exhibiting one of the highest activities. Here, we demonstrate via the use of in vitro trans- and cis-splicing assays that the RNA chaperone activity of L1 is conserved in all three domains of life. However, thermophilic archaeal L1 proteins do not display RNA chaperone activity under the experimental conditions tested here. Furthermore, L1 does not exhibit RNA chaperone activity when in complexes with its cognate rRNA or mRNA substrates. The evolutionary conservation of the RNA chaperone activity among L1 proteins suggests a functional requirement during ribosome assembly, at least in bacteria, mesophilic archaea and eukarya. Surprisingly, rather than facilitating catalysis, the thermophilic archaeal L1 protein from Methanococcus jannaschii (MjaL1) completely inhibits splicing of the group I thymidylate synthase intron from phage T4. Mutational analysis of MjaL1 excludes the possibility that the inhibitory effect is due to stronger RNA binding. To our knowledge, MjaL1 is the first example of a protein that inhibits group I intron splicing.
RNA伴侣活性被定义为蛋白质防止RNA错误折叠或打开错误折叠的RNA构象的能力。大肠杆菌所有大核糖体亚基蛋白的三分之一表现出这种活性,其中L1的活性最高。在这里,我们通过体外转剪接和顺式剪接试验证明,L1的RNA伴侣活性在生命的三个域中都是保守的。然而,嗜热古菌的L1蛋白在此处测试的实验条件下不表现出RNA伴侣活性。此外,L1与其同源rRNA或mRNA底物形成复合物时不表现出RNA伴侣活性。L1蛋白之间RNA伴侣活性的进化保守性表明,至少在细菌、嗜温古菌和真核生物中,核糖体组装过程中有功能需求。令人惊讶的是,来自詹氏甲烷球菌的嗜热古菌L1蛋白(MjaL1)不是促进催化,而是完全抑制噬菌体T4的I组胸苷酸合酶内含子的剪接。对MjaL1的突变分析排除了抑制作用是由于更强的RNA结合的可能性。据我们所知,MjaL1是抑制I组内含子剪接的蛋白质的第一个例子。