Sektion Biowissenschaften der Martin-Luther-Universität Halle-Wittenberg, Neuwerk 1, DDR-4020, Halle, German Democratic Republic.
Planta. 1988 Nov;176(1):51-9. doi: 10.1007/BF00392479.
Compartmentation of uridine 5'-triphosphate (UTP) was studied during the nucleolar synthesis of cytoplasmic ribosomal RNA (cyt-rRNA) and the synthesis of cytoplasmic transfer RNA (cyt-tRNA) in the nuclear matrix as well as the synthesis of mitochondrial ribosomal RNA (mt-rRNA) in tomato (Lycopersicon esculentum Mill. cv. Lukullus) cell-suspension culture using the approach of Wiegers et al. (Eur. J. Biochem. 64, 535-540, 1976). Before measurements were made, it was ensured that: (i) there was steady-state labeling of all RNAs studied as well as UTP; (ii) there was stability of cyt-tRNA and cyt-rRNA; (iii) there was no label randomization through degradation of [(3)H]uridine; (iv) there were significant differences in the specific radioactivity of UTP, the final immediate precursor of RNA, after supplying the cells with two different exogenous [(3)H]uridine concentrations.By comparing the steady-state specific radioactivity of UTP with that of cyt-tRNA and cyt-18S rRNA during constant [(3)H]uridine supply, we found that the three molecules had equal specific radioactivities which, however, differed significantly from that of the mt-rRNA. With a 20-fold higher uridine concentration, i.e. a 20-fold lower specific radioactivity of exogenous [(3)H]uridine, the specific radioactivity of cyt-rRNA, cyt-tRNA and UTP decreased proportionally whereas that of mt-RNA increased. These results argue against different UTP pools during synthesis of cyt-rRNA and cyt-tRNA, but indicate compartmentation of UTP during rRNA synthesis in the nucleus and the mitochondria of tomato cells.
采用 Wiegers 等人(Eur. J. Biochem. 64, 535-540, 1976)的方法,研究了细胞质核糖体 RNA(cyt-rRNA)核仁合成以及细胞质转移 RNA(cyt-tRNA)和线粒体核糖体 RNA(mt-rRNA)合成过程中尿嘧啶 5'-三磷酸(UTP)的分隔情况。在进行测量之前,确保了:(i)所有研究的 RNA 以及 UTP 都处于稳定标记状态;(ii)cyt-tRNA 和 cyt-rRNA 稳定;(iii)通过降解 [(3)H]尿嘧啶,不会使标记随机化;(iv)在向细胞提供两种不同外源性 [(3)H]尿嘧啶浓度后,RNA 的最终直接前体 UTP 的特异性放射性有显著差异。通过比较恒定 [(3)H]尿嘧啶供应期间 UTP 与 cyt-tRNA 和 cyt-18S rRNA 的稳态特异性放射性,我们发现这三种分子具有相同的特异性放射性,但与 mt-rRNA 的特异性放射性有显著差异。当尿嘧啶浓度提高 20 倍,即外源性 [(3)H]尿嘧啶的特异性放射性降低 20 倍时,cyt-rRNA、cyt-tRNA 和 UTP 的特异性放射性成比例下降,而 mt-RNA 的特异性放射性增加。这些结果表明在 cyt-rRNA 和 cyt-tRNA 合成过程中,UTP 没有不同的池,但表明 UTP 在细胞核和线粒体中 rRNA 合成时被分隔。