Sturani E, Costantini M G, Martegani E, Alberghina L
Eur J Biochem. 1979 Aug 15;99(1):1-7. doi: 10.1111/j.1432-1033.1979.tb13224.x.
Mycelia of Neurospora crassa in a steady state of growth in different media have a ribosomal content proportional to the rate of growth. Moreover, both the percentage of polysomes and the average ribosomal activity are about the same at all different growth rates. The content of polyadenylated RNA was determined in three different conditions of exponential growth, which allowed growth rates that ranged from 0.26 to 0.51 duplications/h, and was found to constitute about the same fraction of total RNA (4.5--5.2%). Using a kinetic approach, an equation was derived which allowed determination of the average half-lives of polyadenylated RNA: in each medium the cultures were labeled from the moment of the inoculation with [32P]orthophosphate and were then given a 10-min pulse with [5-3H]uridine when they were in the exponential phase. It was found that the determined half-lives of polyadenylated RNA vary, depending on the growth medium, between 30 and 60 min, but with no direct correlation with the growth rate. Moreover, the rate of synthesis of polyadenylated RNA relative to that of stable RNA decreased with the growth rate. On the basis of previous data on the rates of synthesis of stable RNA, it was possible to make an evaluation of the absolute rate of synthesis of polyadenylated RNA. Whereas the rate of synthesis of stable ribosomal RNA increases as a function of the square of the number of duplications per hour, the increase in the rate of synthesis of polyadenylated RNA with the growth rate is much less consistent. It is concluded that in Neurospora the growth rate does not depend on the rate of synthesis of mRNA but rather on the rate of synthesis of rRNA, which sets both the ribosomal level and the steady-state level of mRNA.
在不同培养基中处于稳定生长状态的粗糙脉孢菌菌丝体,其核糖体含量与生长速率成正比。此外,在所有不同生长速率下,多核糖体的百分比和平均核糖体活性大致相同。在三种不同的指数生长条件下测定了聚腺苷酸化RNA的含量,这些条件下的生长速率范围为0.26至0.51次复制/小时,结果发现聚腺苷酸化RNA占总RNA的比例大致相同(4.5 - 5.2%)。采用动力学方法推导了一个方程,用于确定聚腺苷酸化RNA的平均半衰期:在每种培养基中,培养物从接种时开始用[32P]正磷酸盐标记,然后在指数生长期给予[5-3H]尿苷10分钟的脉冲标记。结果发现,聚腺苷酸化RNA的测定半衰期因生长培养基而异,在30至60分钟之间,但与生长速率没有直接相关性。此外,聚腺苷酸化RNA相对于稳定RNA的合成速率随生长速率而降低。根据先前关于稳定RNA合成速率的数据,可以评估聚腺苷酸化RNA的绝对合成速率。稳定核糖体RNA的合成速率随每小时复制数的平方而增加,而聚腺苷酸化RNA合成速率随生长速率的增加则不太一致。得出的结论是,在粗糙脉孢菌中,生长速率不取决于mRNA的合成速率,而是取决于rRNA的合成速率,rRNA的合成速率既决定了核糖体水平,也决定了mRNA的稳态水平。