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海胆胚胎中的组蛋白基因转换。晚期胚胎组蛋白信使核糖核酸的鉴定及其合成调控。

Histone gene switch in the sea urchin embryo. Identification of late embryonic histone messenger ribonucleic acids and the control of their synthesis.

作者信息

Hieter P A, Hendricks M B, Hemminki K, Weinberg E S

出版信息

Biochemistry. 1979 Jun 26;18(13):2707-16. doi: 10.1021/bi00580a004.

Abstract

During embryogenesis in the sea urchin Strongylocentrotus purpuratus, there is a shift from one histone mRNA population to another. The early and late embryonic histone mRNAs, previously shown to differ considerably in sequence from each other by hybrid melting studies, are shown here to differ also in electrophoretic mobility on polyacrylamide gels as the positions of the early and late mRNAs are completely noncoincident. The various species of both early and late samples are identified as particular histone mRNAs by hybridization to cloned histone DNAs containing part of the early-type repeat unit or to restriction enzyme fragments derived from these unit. Four bands in the early mRNA sample are identified as H1, H3, H2A " H2B, and H4 mRNA while at least 10 bands can be seen in the late mRNA preparation with unambiguous identification of H1, H2B, and H4 mRNAs. A cluster of late species is shown to contain both H3 and H2A mRNA. When a polysomal RNA preparation from the 26-h embryo is hybridized to the histone DNA, eluted, and then translated in vitro in a wheat germ system, the histone products migrate in the position of late histones when subjected to electrophoresis on Triton X-urea gels. Using DNA which contains genes for H2A + H3 or H2A alone, we demonstrate the specificity of the early-type DNA probes for these two late histones. Therefore, by hybridization of newly synthesized RNAs and translation of the total polysomal RNA present in the late embryo, it is shown that mRNAs for all five histone classes may cross-react with the cloned early-type DNA. The hybrids formed, however, are much less stable than those formed with the early histone mRNA. In vitro translation of total cytoplasmic RNA from various embryonic stages indicates that transition between the two classes occurs during most of the blastula period.

摘要

在紫海胆胚胎发生过程中,会从一种组蛋白mRNA群体转变为另一种。早期和晚期胚胎组蛋白mRNA,此前通过杂交熔解研究已表明它们在序列上彼此有很大差异,此处通过聚丙烯酰胺凝胶电泳迁移率也显示出差异,因为早期和晚期mRNA的位置完全不重合。通过与包含部分早期型重复单元的克隆组蛋白DNA或从这些单元衍生的限制性酶切片段杂交,早期和晚期样品的各种种类被鉴定为特定的组蛋白mRNA。早期mRNA样品中的四条带被鉴定为H1、H3、H2A、H2B和H4 mRNA,而在晚期mRNA制剂中可以看到至少十条带,其中H1、H2B和H4 mRNA得到明确鉴定。一组晚期种类显示同时含有H3和H2A mRNA。当将来自26小时胚胎的多聚核糖体RNA制剂与组蛋白DNA杂交、洗脱,然后在小麦胚芽系统中进行体外翻译时,组蛋白产物在Triton X - 尿素凝胶上电泳时迁移到晚期组蛋白的位置。使用含有H2A + H3或单独H2A基因的DNA,我们证明了早期型DNA探针对于这两种晚期组蛋白的特异性。因此,通过新合成RNA的杂交以及晚期胚胎中存在的总多聚核糖体RNA的翻译,表明所有五种组蛋白类别的mRNA都可能与克隆的早期型DNA发生交叉反应。然而,形成的杂交体比与早期组蛋白mRNA形成的杂交体稳定性要低得多。来自不同胚胎阶段的总细胞质RNA的体外翻译表明,这两类之间的转变发生在囊胚期的大部分时间。

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