Steege D A, Graves M C, Spremulli L L
J Biol Chem. 1982 Sep 10;257(17):10430-9.
RNA sequencing methods have been used to determine sequences at the 3' ends of the small subunit RNAs from Euglena gracilis cytoplasmic and chloroplast ribosomes. The cytoplasmic rRNA sequence, AUCAU psi GOH, has features typical of eukaryotic 18 S rRNAs. The sequence at the 3' terminus of the chloroplast 17 S rRNA, AACAACUCCCOH, differs in several positions from nucleotides conserved in both prokaryotic and eukaryotic small subunit rRNAs, but it does terminate with a pyrimidine tract. Like prokaryotic ribosomes, the chloroplast ribosome displays highly specific binding to purine-rich oligonucleotides. The 30 S subunits select from a random mixture of oligonucleotides only those that contain the sequence GGGAG, which is complementary to the 3' end of 17 S rRNA. The 68S chloroplast ribosome has been shown to be a substrate in vitro for colicin E3. Colicin E3 introduces a cleavage at a position in the chloroplast 17 S rRNA that corresponds exactly to that hydrolyzed in Escherichia coli S rRNA. These results suggest that the 3'-terminal region of 17 S rRNA has an arrangement in the 68 S chloroplast ribosome that may be equivalent to that of the prokaryotic ribosome, with the 3' end exposed in the 30 S subunit for interactions with complementary RNA sequences.
RNA测序方法已被用于确定纤细裸藻细胞质和叶绿体核糖体小亚基RNA 3'末端的序列。细胞质rRNA序列AUCAUψGOH具有真核生物18S rRNA的典型特征。叶绿体17S rRNA 3'末端的序列AACAACUCCCOH在几个位置上与原核生物和真核生物小亚基rRNA中保守的核苷酸不同,但它确实以嘧啶序列结尾。与原核生物核糖体一样,叶绿体核糖体对富含嘌呤的寡核苷酸表现出高度特异性结合。30S亚基从寡核苷酸的随机混合物中仅选择那些包含与17S rRNA 3'末端互补的序列GGGAG的寡核苷酸。68S叶绿体核糖体已被证明是体外大肠杆菌素E3的底物。大肠杆菌素E3在叶绿体17S rRNA中的一个位置引入切割,该位置与在大肠杆菌S rRNA中水解的位置完全对应。这些结果表明,17S rRNA的3'末端区域在68S叶绿体核糖体中的排列可能与原核生物核糖体的排列相同,3'末端暴露在30S亚基中以与互补RNA序列相互作用。