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Regulation of ribosomal protein gene expression in higher eukaryotes

高等真核生物核糖体蛋白基因表达的调控

基本信息

批准号：
01570121
负责人：
TANAKA Tatsuo
金额：
$ 1.41万
依托单位：
University of the Ryukyus (1990-1991)Yamagata University (1989)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

(1)Complementary DNA cloning.We isolated CDNA clones for two rat ribososomal proteins and four chicken ribosomal proteins. The CDNAs were not only used as probes in gene cloning but also sequenced to provide information on the structure, function and evolution of the ribosomes.(2)Gene cloning.We isolated the clones of four kinds of ribosomal proteins genes, L5, L7a, L30, and L37a, from a chicken genomic library and determined the entire nucleotide sequences of the genes, expanding about 8.5, 4.5, 4.0, and 3.5 kilobases, respectively.(3)Features in the promoter region of the ribosomal protein genes.These genes do not have either canonical CAAT box or TATA box. There are many CpG dimers concentrated around the transcription initiation site and several GC boxes, known as the binding sites of the transcription factor Spl, in the promoter region. These findings are common to many other house keeping genes.We could not found specific nucleotide sequence that is common to promoter regions of all four genes and seems to be concerned in transcriptional regulation. But there are several candidates for the binding subs of the transcriptional regulation factors and/or initiation factors common to two or three genes. The sequence similar to the internal transcriptional control region of 5S RNA gene was also found in the promoter region of the gene of L5, the 5S RNA binding protein.These findings suggest that the expression of ribosomal protein gene is regulated by complicated control mechanisms. The function of these candidates of regulatory elements is being analyzed by CAT assay and the binding assay of nuclear extract.

(1)互补DNA克隆。我们分离了两种大鼠核糖体蛋白和四种鸡核糖体蛋白的cDNA克隆。 cDNA不仅可以作为基因克隆的探针，还可以通过测序提供有关核糖体结构、功能和进化的信息。(2)基因克隆。我们从鸡基因组文库中分离出了L5、L7a、L30和L37a四种核糖体蛋白基因的克隆，并确定了基因的完整核苷酸序列，扩增了约8.5、4.5、分别为4.0和3.5 kilobase。(3)核糖体蛋白基因启动子区域的特征。这些基因既没有典型的CAAT盒，也没有TATA盒。转录起始位点周围集中有许多 CpG 二聚体，启动子区域有几个 GC 盒（称为转录因子 Spl 的结合位点）。这些发现对于许多其他管家基因来说是常见的。我们无法找到所有四个基因的启动子区域共有的特定核苷酸序列，并且似乎与转录调控有关。但是对于两个或三个基因共有的转录调节因子和/或起始因子的结合子有几种候选者。在5S RNA结合蛋白L5基因的启动子区也发现了与5S RNA基因内部转录控制区相似的序列。这些发现表明核糖体蛋白基因的表达受到复杂的控制机制的调节。正在通过 CAT 测定和核提取物结合测定来分析这些候选调控元件的功能。