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鹌鹑输卵管中年龄依赖性基因诱导。十一。转录后水平的改变(分析方面)。

Age-dependent gene induction in quail oviduct. XI. Alterations on the post-transcriptional level (analytical aspect).

作者信息

Arendes J, Zahn R K, Müller W E

出版信息

Mech Ageing Dev. 1980 Sep-Oct;14(1-2):49-57. doi: 10.1016/0047-6374(80)90105-0.

Abstract

For the elucidation of the age-dependent reduction of avidin synthesis in the oviduct of quails studies on on the level of post-transcriptional modification of mRNA were performed. Incorporation of [3H] adenosine into mRNA is drastically reduced in senescent animals compared to adult animals. Double labeling of mRNA with [3H]-adenosine and [3H] uridine revealed that in the presence of actinomycin D the incorporation ratio adenosine/uridine into mRNA increases to higher values in the case of adult animals compared with the ratio determined for mRNA from senescent animals. These data support the assumption that in senescent animals the length of the poly(A) sequence on mRNA might be shorter. Because of the low efficiency of label incorporation into poly(A) in whole animals, a method for directly introducing a radioactive label in vitro into the isolated poly(A) was developed for the estimation of the length of the poly(A) segments. A randomly labeled product with no apparent degradation is achieved by methylation of poly(A) with [3H] dimethyl sulphate. Therefore the method is a powerful tool for the detection and size determination of small amounts of poly(A). Size determination of poly(A) from oviducts of adult and senescent quails by the methylation method revealed that in the case of senescent animals an increase in shorter poly(A) tracts can be detected. These results support the assumption that the age-dependent reduction of avidin synthesis is also caused by changes in the control mechanisms on the post-transcriptional level.

摘要

为了阐明鹌鹑输卵管中抗生物素蛋白合成随年龄增长而减少的现象,我们对mRNA转录后修饰水平进行了研究。与成年动物相比,衰老动物中[3H]腺苷掺入mRNA的量大幅减少。用[3H] - 腺苷和[3H]尿苷对mRNA进行双重标记显示,在放线菌素D存在的情况下,成年动物mRNA中腺苷/尿苷的掺入率相对于衰老动物mRNA的掺入率会增加到更高的值。这些数据支持了衰老动物中mRNA上多聚腺苷酸(poly(A))序列长度可能较短的假设。由于在完整动物中标记掺入多聚腺苷酸的效率较低,我们开发了一种在体外将放射性标记直接引入分离的多聚腺苷酸中的方法,用于估计多聚腺苷酸片段的长度。用[3H]硫酸二甲酯对多聚腺苷酸进行甲基化可得到无明显降解的随机标记产物。因此,该方法是检测和测定少量多聚腺苷酸大小的有力工具。通过甲基化方法对成年和衰老鹌鹑输卵管中的多聚腺苷酸进行大小测定表明,在衰老动物中可以检测到较短多聚腺苷酸片段的增加。这些结果支持了抗生物素蛋白合成随年龄增长而减少也是由转录后水平控制机制变化引起的这一假设。

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