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生长素调节的基因表达。

Auxin-regulated gene expression.

作者信息

Key J L, Kroner P, Walker J, Hong J C, Ulrich T H, Ainley W M, Gantt J S, Nagao R T

出版信息

Philos Trans R Soc Lond B Biol Sci. 1986 Nov 17;314(1166):427-40. doi: 10.1098/rstb.1986.0063.

Abstract

During the 1960s a wide range of studies provided an information base that led to the suggestion that auxin-regulated cell processes--especially cell elongation--may be mediated by auxin-regulated gene expression. Indirect evidence from our work, based on the influence of inhibitors of RNA synthesis (e.g. actinomycin D) and of protein synthesis (e.g. cycloheximide) on auxin-induced cell elongation, coupled with correlations of the influence of auxin on RNA synthesis and cell elongation, provided the basis for this suggestion. With the availability of techniques for DNA-DNA and DNA-RNA hybridization, mRNA isolation-translation, in vitro 2D gel analysis of the translation products, and ultimately the cloning by recombinant DNA technologies of genomic DNA and copy DNAs (cDNAs) made to poly(A)+ mRNAs, we and others have provided direct evidence for the influence of auxin on the expression of a few genes (i.e. poly(A)+ RNA levels). Our laboratory has provided evidence for auxin's both down-regulating and up-regulating the level of a few poly(A)+ mRNAs out of a population of about 4 X 10(4) sequences that are not significantly affected by auxin. In our studies on auxin-regulated cell elongation, two cDNA clones (pJCW1 and pJCW2) were isolated which corresponded to poly(A)+ mRNAs that responded during growth transitions in a way consistent with a potential role of their protein products in cell elongation. These mRNAs are most abundant in the elongating zone of the soybean hypocotyl. Upon excision and incubation in the absence of auxin, these mRNAs deplete in concert with a decreasing rate of cell elongation. Addition of auxin to the medium results in both increased levels of these mRNAs and enhanced rates of cell elongation. These mRNAs do not deplete if auxin is added to the medium at the onset of excised incubation, and cell elongation rates remain high. We have isolated and sequenced genomic clones that are homologous to these cDNAs. Of the two genes sequenced, both genes are members of small multigene families. There are regions of high amino acid homology even though the nucleotide sequences are sufficiently different in these regions for cross-hybridization of the clones not to be observed. More recently others, especially Guilfoyle's laboratory, have shown that auxin selectively and rapidly influences the level of certain mRNAs and proteins. We have worked on other gene systems such as ribosomal proteins and possible cell wall proteins that are responsive to auxin; again the nature of regulation of expression of these genes is not known.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

在20世纪60年代,一系列广泛的研究提供了一个信息库,由此有人提出,生长素调节的细胞过程——尤其是细胞伸长——可能是由生长素调节的基因表达介导的。我们的研究提供了间接证据,基于RNA合成抑制剂(如放线菌素D)和蛋白质合成抑制剂(如环己酰亚胺)对生长素诱导的细胞伸长的影响,再加上生长素对RNA合成和细胞伸长影响的相关性,为这一观点提供了依据。随着DNA-DNA和DNA-RNA杂交技术、mRNA分离-翻译技术、翻译产物的体外二维凝胶分析技术的出现,以及最终通过重组DNA技术克隆基因组DNA和针对多聚腺苷酸加尾mRNA合成的互补DNA(cDNA),我们和其他人提供了生长素对少数基因表达(即多聚腺苷酸加尾RNA水平)有影响的直接证据。我们实验室已经证明,在大约4×10⁴个不受生长素显著影响的序列群体中,生长素既能下调也能上调少数多聚腺苷酸加尾mRNA的水平。在我们关于生长素调节细胞伸长的研究中,分离出了两个cDNA克隆(pJCW1和pJCW2),它们对应于在生长转变过程中发生响应的多聚腺苷酸加尾mRNA,其响应方式与其蛋白质产物在细胞伸长中的潜在作用一致。这些mRNA在大豆下胚轴的伸长区最为丰富。在切除并在无生长素的条件下培养时,这些mRNA会随着细胞伸长速率的降低而协同减少。向培养基中添加生长素会导致这些mRNA水平升高以及细胞伸长速率加快。如果在切除培养开始时向培养基中添加生长素,这些mRNA不会减少,细胞伸长速率也会保持较高。我们已经分离并测序了与这些cDNA同源的基因组克隆。在测序的两个基因中,这两个基因都是小多基因家族的成员。尽管这些区域的核苷酸序列差异足够大,以至于未观察到克隆之间的交叉杂交,但仍存在高度的氨基酸同源区域。最近,其他人,尤其是吉尔福伊尔的实验室,已经表明生长素能选择性且迅速地影响某些mRNA和蛋白质的水平。我们研究了其他对生长素作出响应的基因系统,如核糖体蛋白和可能的细胞壁蛋白;同样,这些基因表达的调控性质尚不清楚。

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