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海胆胚胎中多聚核糖体mRNA的合成与周转

Synthesis and turnover of polysomal mRNAs in sea urchin embryos.

作者信息

Galau G A, Lipson E D, Britten R J, Davidson E H

出版信息

Cell. 1977 Mar;10(3):415-32. doi: 10.1016/0092-8674(77)90029-0.

Abstract

The synthesis and turnover kinetics of polysomal mRNA have been measured in sea urchin embryos. Polysomes were isolated from stages ranging between mesenchyme blastula and late gastrula Strongylocentrotus purpuratus embryos which had been exposed to exogenous 3H-guanosine. The amount of radioactivity incorporated into messenger and ribosomal RNAs was determined separately as a function of time, and the precursor pool specific activity was measured in the same embryos. Synthesis and decay rate constants were extracted from the data by a least-squares procedure. Per embryo, the rate of mRNA synthesis was calculated to be about 0.13 pg min-1, while the rate of rRNA synthesis is about 0.022 pg min-1. The newly synthesized mRNA turns over with a half-time of 5.7 hr. The data support only a single decay rate for the mRNA, but small fractions of mRNA decaying at different rates cannot be excluded. Previous studies have shown that a minor fraction of the mRNA includes the least abundant, most highly diverse set of messages ("complex class" mRNAs). To determine whether mRNAs of the complex class are synthesized and degraded at similar rates, labeled mRNA was measured in hybrids formed in mRNA excess reactions with single copy DNA. These experiments showed that complex class mRNAs represent an approximately proportional amount of the new mRNA symthesis, and turn over at the same average rate as does the bulk of the mRNA. Most of the mRNAs in the embryo polysomes are newly synthesized, rather than maternal. This statement refers both to complex class mRNAs and to prevalent mRNAs. Considering the sequence homology between embryo and oocyte mRNAs shown earlier, these results indicate that many of the same structural genes active during oogenesis are being transcribed in embryos at these stages.

摘要

已对海胆胚胎中多聚核糖体mRNA的合成及周转动力学进行了测定。从暴露于外源性3H-鸟苷的紫海胆胚胎的间充质囊胚期至原肠胚后期的各个阶段分离出多聚核糖体。分别测定信使RNA和核糖体RNA中掺入的放射性活度随时间的变化情况,并在相同胚胎中测量前体库的比活度。通过最小二乘法从数据中提取合成和衰变速率常数。每个胚胎的mRNA合成速率经计算约为0.13 pg min-1,而rRNA合成速率约为0.022 pg min-1。新合成的mRNA的半衰期为5.7小时。这些数据仅支持mRNA的单一衰变速率,但不能排除一小部分mRNA以不同速率衰变的可能性。先前的研究表明,一小部分mRNA包括丰度最低、多样性最高的一组信息(“复杂类”mRNA)。为了确定复杂类mRNA的合成和降解速率是否相似,在与单拷贝DNA的mRNA过量反应中形成的杂交体中测量了标记的mRNA。这些实验表明,复杂类mRNA在新mRNA合成中所占比例大致相当,并且与大部分mRNA以相同的平均速率周转。胚胎多聚核糖体中的大多数mRNA是新合成的,而非母源性的。这一说法既适用于复杂类mRNA,也适用于普遍存在的mRNA。鉴于先前显示的胚胎mRNA与卵母细胞mRNA之间的序列同源性,这些结果表明,许多在卵子发生过程中活跃的相同结构基因在这些阶段的胚胎中也在转录。

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