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DNA甲基化是悬铃木(Acer pseudoplatanus L.)液体培养细胞的造粉体中光合作用基因表达的决定性因素。

DNA methylation is a determinative element of photosynthesis gene expression in amyloplasts from liquid-cultured cells of sycamore (Acer pseudoplatanus L.).

作者信息

Ngernprasirtsiri J, Kobayashi H, Akazawa T

机构信息

Research Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Japan.

出版信息

Cell Struct Funct. 1990 Oct;15(5):285-93. doi: 10.1247/csf.15.285.

Abstract

Transcriptional regulation has been shown to operate as a selective control mechanism of expression of photosynthetic genes in the nonphotosynthetic plastids, amyloplasts, of a white-wild cell line of sycamore (Acer pseudoplatanus L.). To elaborate the mechanisms governing the transcriptional regulation at the molecular level, we have examined the template activity of the amyloplast DNA compared to the chloroplast DNA by using the in vitro run-off transcription assay system with extracts of the two plastid types. The results of these assays clearly indicate that most of the amyloplast DNA regions do not serve as a template for the in vitro transcription regardless of the plastid extracts; this is in contrast to the chloroplast DNA which serves as an active template. It is highly likely that the template activity of amyloplast DNA per se is the modulating element of transcriptional regulation. Parallel experiments determining the DNA base content by HPLC analysis have shown that a variety of methylated bases, especially 5-methylcytosine, are localized in the DNA regions containing suppressed genes of the amyloplast genome. In sharp contrast, methylated bases were undetectable in the expressed gene regions of amyloplast and whole chloroplast genomes. The overall findings strongly support the notion that DNA methylation is involved in the selective suppression of photosynthetic genes in the nonphotosynthetic plastids of cultured sycamore cells.

摘要

转录调控已被证明是一种选择性控制机制,作用于悬铃木(Acer pseudoplatanus L.)白色野生细胞系中非光合质体——造粉体中光合基因的表达。为了在分子水平上详细阐述转录调控的机制,我们通过使用来自两种质体类型提取物的体外径流转录分析系统,比较了造粉体DNA与叶绿体DNA的模板活性。这些分析结果清楚地表明,无论使用哪种质体提取物,大多数造粉体DNA区域都不能作为体外转录的模板;这与作为活性模板的叶绿体DNA形成对比。造粉体DNA本身的模板活性很可能是转录调控的调节因素。通过HPLC分析确定DNA碱基含量的平行实验表明,多种甲基化碱基,尤其是5 - 甲基胞嘧啶,定位于造粉体基因组中含有被抑制基因的DNA区域。与之形成鲜明对比的是,在造粉体和整个叶绿体基因组的表达基因区域中未检测到甲基化碱基。总体研究结果有力地支持了这样一种观点,即DNA甲基化参与了培养的悬铃木细胞中非光合质体中光合基因的选择性抑制。

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