Beltrán-Peña E, Ortíz-López A, Sánchez de Jiménez E
Departamento de Bioquímica, Facultad de Química, UNAM, México, D.F., México.
Plant Mol Biol. 1995 May;28(2):327-36. doi: 10.1007/BF00020251.
In several eukaryotic organisms the mRNA expression for ribosomal proteins (RPs) is highly regulated at the translational level. The present work addresses the synthesis of RPs early in germination and particularly the origin of their transcripts. Excised maize axes were labeled with [35S]methionine for 6 h, and both cytoplasmic and ribosomal proteins were analyzed by electrophoresis and fluorography. It was found that some RPs are newly synthesized and already incorporated into ribosomes during this period. Synthesis of RPs was also observed under conditions of transcription inhibition, suggesting the presence of stored-RP transcripts in the embryonic axes. In vitro translation of polysomes isolated from alpha-amanitin-treated axes early in germination (3 and 9 h) was performed. Immunoprecipitation of the products revealed the synthesis of rps. Four heterologous probes two each of the small subunit (S4 and S6) and large (L3, L16) subunit rps were used to detect the correspondent transcripts within the stored mRNA stock of the embryonic axes. Both, by slot blot and northern analysis, a positive cross-reaction occurred for all the tested samples. Although only two of them (S4 and S6) seem to be stored as mature mRNA.
在几种真核生物中,核糖体蛋白(RPs)的mRNA表达在翻译水平上受到高度调控。目前的工作研究了萌发早期核糖体蛋白的合成,特别是其转录本的来源。将切除的玉米胚轴用[35S]甲硫氨酸标记6小时,通过电泳和荧光自显影分析细胞质蛋白和核糖体蛋白。结果发现,在此期间一些核糖体蛋白是新合成的并且已经整合到核糖体中。在转录抑制条件下也观察到核糖体蛋白的合成,这表明胚轴中存在储存的核糖体蛋白转录本。对萌发早期(3小时和9小时)用α-鹅膏蕈碱处理的胚轴分离的多核糖体进行体外翻译。产物的免疫沉淀显示核糖体蛋白的合成。使用四个异源探针,小亚基(S4和S6)和大亚基(L3,L16)的核糖体蛋白各两个,来检测胚轴储存mRNA库中的相应转录本。通过狭缝印迹和Northern分析,所有测试样品均出现阳性交叉反应。尽管其中只有两个(S4和S6)似乎以成熟mRNA的形式储存。