Patel C V, Gopinathan K P
Microbiology and Cell Biology Department, Indian Institute of Science, Bangalore.
Indian J Biochem Biophys. 1991 Oct-Dec;28(5-6):521-30.
The contents of fibroin H RNA as a function of development have been quantitated in the posterior silk glands of Bombyx mori larvae on different days of 4th and 5th instars. The fibroin RNA levels increased during the feeding stages of larvae and the RNA got completely degraded during the interim moult. The patterns of accumulation of fibroin RNA were similar in both the instars. Although there was considerable increase in the fibroin RNA content during the 5th larval instar, the relative abundance of fibroin RNA in the total RNA was fairly constant during the 4th and 5th instars. The increased content of fibroin RNA in 5th instar was the consequence of an overall increase in transcription accompanying the development progress, rather than specific increase only in fibroin transcription. The contents of fibroin protein in the 4th and 5th instars of development have also been quantitated making use of a sensitive radioimmune assay with a purified, antifibroin antibody. There were substantial differences between 4th and 5th instars in the absolute fibroin contents as well as the relative proportion of fibroin in the total proteins. These results implied that although the fibroin gene was transcribed at the same efficiency during the 4th and 5th instars, the translational efficiency was much lower during the 4th instar. The extent of polyadenylation of fibroin RNA was similar in both instars. However, there was a two-fold increase in the polysome association of fibroin RNA in the 5th instar. Over and above this, there was substantial increase during the 5th instar in the contents of those tRNAs. (e.g. Gly, Ala and Ser) which are abundantly represented in fibroin and therefore directly related to the expression of fibroin. The increased polysome association of fibroin mRNA and the adequate supply of cognate tRNAs in the 5th instar, together contributes to the translational regulation of fibroin in a developmental stage-specific manner. Based on these observations, we propose that translational regulation plays a major role in the development stage-specific synthesis of fibroin in Bombyx mori.
在家蚕幼虫四龄和五龄不同天数的后部丝腺中,已对丝心蛋白H RNA含量随发育阶段的变化进行了定量分析。丝心蛋白RNA水平在幼虫取食阶段升高,而在中间蜕皮期RNA完全降解。两个龄期丝心蛋白RNA的积累模式相似。尽管在五龄幼虫期丝心蛋白RNA含量有显著增加,但在四龄和五龄期间,丝心蛋白RNA在总RNA中的相对丰度相当恒定。五龄期丝心蛋白RNA含量的增加是伴随发育进程转录总体增加的结果,而非仅丝心蛋白转录的特异性增加。利用纯化的抗丝心蛋白抗体通过灵敏的放射免疫测定法,也对发育四龄和五龄期的丝心蛋白含量进行了定量。四龄和五龄期在丝心蛋白绝对含量以及丝心蛋白在总蛋白中的相对比例方面存在显著差异。这些结果表明,尽管丝心蛋白基因在四龄和五龄期以相同效率转录,但四龄期的翻译效率要低得多。两个龄期丝心蛋白RNA的聚腺苷酸化程度相似。然而,五龄期丝心蛋白RNA与多核糖体的结合增加了两倍。除此之外,五龄期那些在丝心蛋白中大量存在且因此与丝心蛋白表达直接相关的tRNA(如甘氨酸、丙氨酸和丝氨酸tRNA)含量大幅增加。五龄期丝心蛋白mRNA与多核糖体结合的增加以及同源tRNA的充足供应,共同以发育阶段特异性方式促进了丝心蛋白的翻译调控。基于这些观察结果,我们提出翻译调控在家蚕丝心蛋白发育阶段特异性合成中起主要作用。