Iio T, Tamaoki T
Can J Biochem. 1976 May;54(5):408-12. doi: 10.1139/o76-059.
Intracellular distribution of active mRNAs for alpha-fetoprotein (alphaFP) and albumin in fetal and adult mouse liver was studied. Livers were fractionated into nucleus and cytoplasm. The latter was further fractionated by sucrose density-gradient centrifugation into four subfractions, the top fraction containing soluble components, the 1.0 M sucrose layer containing primarily 80S ribosomes, the 1.5 M sucrose layer containing light polysomes and the pellets containing heavy polysomes. RNA was extracted from each fraction and its ability to direct the synthesis of alphaFP and albumin was determined in a mouse sarcoma 180 cell-free system. Distribution of alphaFP and albumin mRNAs in fetal mouse liver was similar; 2% in the nucleus, 98% in the cytoplasm, of which more than 90% was found in the polysome fractions. The results suggest that alphaFP and albumin mRNAs, once formed, are quickly and efficiently utilized for protein synthesis. The major proportion of albumin mRNA in adult mouse liver was also found to be associated with polysomes. However, the amount of translatable alphaFP mRNA was low in all subcellular fractions examined, suggesting that transcription or processing of alphaFP mRNA is defective in adult mouse liver.
研究了胎鼠和成年小鼠肝脏中甲胎蛋白(αFP)和白蛋白活性mRNA的细胞内分布。将肝脏分离为细胞核和细胞质。后者通过蔗糖密度梯度离心进一步分离为四个亚组分,顶部组分包含可溶性成分,1.0M蔗糖层主要包含80S核糖体,1.5M蔗糖层包含轻多核糖体,沉淀包含重多核糖体。从每个组分中提取RNA,并在小鼠肉瘤180无细胞系统中测定其指导αFP和白蛋白合成的能力。胎鼠肝脏中αFP和白蛋白mRNA的分布相似;细胞核中占2%,细胞质中占98%,其中90%以上存在于多核糖体组分中。结果表明,αFP和白蛋白mRNA一旦形成,就会迅速有效地用于蛋白质合成。在成年小鼠肝脏中也发现白蛋白mRNA的主要部分与多核糖体相关。然而,在所检查的所有亚细胞组分中,可翻译的αFP mRNA含量都很低,这表明成年小鼠肝脏中αFP mRNA的转录或加工存在缺陷。