Linz J E, Orlowski M
J Bacteriol. 1982 Jun;150(3):1138-44. doi: 10.1128/jb.150.3.1138-1144.1982.
When introduced into nutrient medium under air, the asexual sporangiospores of Mucor racemosus germinated with 5 to 8 h, culminating with the emergence of germ tubes. We found that sporangiospores increased 20% in dry weight during the first 60 min of germination, indicating a high degree of synthetic activity. Sucrose density gradient analysis of spore extracts revealed that the percentage of ribosomes associated with mRNA increased from 22.5% in dormant spores to 85% within 10 min after the addition of medium and remained at this level for at least 3 h. L-[14C]leucine was immediately incorporated at a rapid rate into protein of a leucine auxotroph, whereas [3H]uracil or [32P]phosphate was incorporated into RNA at a significant rate only 20 min after the addition of medium. This newly synthesized RNA occurred in polysomes only after 30 min had passed. Pool synthesized RNA occurred in polysomes only after 30 min had passed. Pool equilibration of the radioactive precursors was not limiting to these measurements. Polyadenylated RNA was isolated from dormant spores by oligodeoxythymidylic acid-cellulose chromatography and was found to comprise 3.3% of the total cellular RNA. Sucrose density gradient centrifugation revealed the polyadenylated RNA to be heterodisperse in size, ranging from 6S to 20S. It was concluded that M. racemosus sporangiospores contain preformed mRNA which is translated commencing immediately upon the addition of nutrient medium.
当置于有氧的营养培养基中时,总状毛霉的无性孢子囊孢子在5至8小时内萌发,最终长出芽管。我们发现,孢子囊孢子在萌发的最初60分钟内干重增加了20%,这表明其具有高度的合成活性。对孢子提取物进行蔗糖密度梯度分析发现,与mRNA结合的核糖体百分比从休眠孢子中的22.5%在添加培养基后10分钟内增加到85%,并在该水平至少维持3小时。L-[14C]亮氨酸立即以快速速率掺入亮氨酸营养缺陷型的蛋白质中,而[3H]尿嘧啶或[32P]磷酸盐仅在添加培养基20分钟后才以显著速率掺入RNA中。这种新合成的RNA仅在30分钟后才出现在多核糖体中。放射性前体的库平衡对这些测量没有限制。通过寡聚脱氧胸苷酸-纤维素色谱法从休眠孢子中分离出聚腺苷酸化RNA,发现其占细胞总RNA的3.3%。蔗糖密度梯度离心显示聚腺苷酸化RNA在大小上是异质分散的,范围从6S到20S。得出的结论是,总状毛霉孢子囊孢子含有预先形成的mRNA,在添加营养培养基后立即开始翻译。