Wosnick M A, White B N
Biochim Biophys Acta. 1979 Jan 26;561(1):194-205. doi: 10.1016/0005-2787(79)90502-1.
A simple procedure for the purification of Drosophila tRNAs which contain the hypermodified nucleoside Q was developed. The cis-diol group of Q renders it susceptible to attack by sodium periodate, with the resultant formation of a hydrophobic pyrrol ring structure. The increase in hydrophobicity after periodate modification, of those tRNA species which contain the Q nucleoside, causes them to be selectively retarded on benzoylated DEAE-cellulose or RPC-5 columns, and allows their isolation in essentially pure form. Utilizing this property, the Q-containing tyrosine tRNA from Drosophila, tRNATyr1delta, was purified. The non-Q-containing tyrosine tRNA, tRNA, tRNATyr1gamma, was also purified, and the nucleoside compositions of the two were determined and compared. In addition to the Q nucleoside, these tRNAs appear to differ by the presence (tRNATyr1delta) or absence (tRNATyr1gamma) of 5-methylcytidine. In all other respects, these tRNAs appear to be identical, and are probably products of the same gene which differ in their levels of post-transcriptional modification.
开发了一种简单的纯化果蝇中含有超修饰核苷Q的tRNA的方法。Q的顺式二醇基团使其易受高碘酸钠攻击，从而形成疏水性吡咯环结构。高碘酸盐修饰后，那些含有Q核苷的tRNA种类的疏水性增加，导致它们在苯甲酰化DEAE-纤维素或RPC-5柱上被选择性阻滞，并允许以基本纯的形式分离它们。利用这一特性，纯化了果蝇中含Q的酪氨酸tRNA，即tRNATyr1δ。不含Q的酪氨酸tRNA，即tRNATyr1γ，也被纯化，并测定和比较了两者的核苷组成。除了Q核苷外，这些tRNA似乎因5-甲基胞苷的存在（tRNATyr1δ）或不存在（tRNATyr1γ）而有所不同。在所有其他方面，这些tRNA似乎是相同的，可能是同一基因的产物，只是转录后修饰水平不同。
