Vila-Sanjurjo Antón, Ridgeway William K, Seymaner Veysel, Zhang Wen, Santoso Steve, Yu Kexin, Cate Jamie H Doudna
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):8682-7. doi: 10.1073/pnas.1133380100. Epub 2003 Jul 9.
Protein biosynthesis on the ribosome requires accurate reading of the genetic code in mRNA. Two conformational rearrangements in the small ribosomal subunit, a closing of the head and body around the incoming tRNA and an RNA helical switch near the mRNA decoding site, have been proposed to select for complementary base-pairing between mRNA codons and tRNA anticodons. We determined x-ray crystal structures of the WT and a hyper-accurate variant of the Escherichia coli ribosome at resolutions of 10 and 9 A, respectively, revealing that formation of the intact 70S ribosome from its two subunits closes the conformation of the head of the small subunit independent of mRNA decoding. Moreover, no change in the conformation of the switch helix is observed in two steps of tRNA discrimination. These 70S ribosome structures indicate that mRNA decoding is coupled primarily to movement of the small subunit body, consistent with previous proposals, whereas closing of the head and the helical switch may function in other steps of protein synthesis.
核糖体上的蛋白质生物合成需要准确读取mRNA中的遗传密码。小核糖体亚基中的两种构象重排,即围绕进入的tRNA头部和主体的闭合以及mRNA解码位点附近的RNA螺旋开关,被认为是为了选择mRNA密码子和tRNA反密码子之间的互补碱基配对。我们分别以10埃和9埃的分辨率确定了野生型和大肠杆菌核糖体的超精确变体的X射线晶体结构,结果表明,由其两个亚基形成完整的70S核糖体可闭合小亚基头部的构象,而与mRNA解码无关。此外,在tRNA识别的两个步骤中未观察到开关螺旋构象的变化。这些70S核糖体结构表明,mRNA解码主要与小亚基主体的运动相关,这与之前的提议一致,而头部和螺旋开关的闭合可能在蛋白质合成的其他步骤中起作用。