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盘基网柄菌营养体特异性基因的一级结构与调控

Primary structure and regulation of vegetative specific genes of Dictyostelium discoideum.

作者信息

Singleton C K, Manning S S, Ken R

机构信息

Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235.

出版信息

Nucleic Acids Res. 1989 Dec 11;17(23):9679-92. doi: 10.1093/nar/17.23.9679.

Abstract

We have examined the expression and structure of several genes belonging to two classes of vegetative specific genes of the simple eukaryote, Dictyostelium discoideum. In amebae grown on bacteria, deactivation of all vegetative specific genes occurred at the onset of development and very little mRNA exists by 8 to 10 hours. In contrast, when cells were grown in axenic broth, the mRNA levels remained constant until a dramatic drop occurred around 10 to 12 hours. Thus, regulation of both classes of genes during the first several hours of development is dependent upon the prior growth conditions. Analysis of genomic clones has resulted in the identification of two V genes, V1 and V18, as ribosomal protein genes. Several other V genes were not found to be ribosomal protein genes, suggesting that in Dictyostelium non-ribosomal protein genes may be coordinately regulated with the ribosomal protein genes. Finally, using deletion analysis we show that the promoters of two of the V genes are composed of a constitutive positive element(s) located upstream of sequences involved in the regulated expression of these genes and within the first 545 upstream bp for V18 and 850 bp for V14. The regions involved in regulated expression were localized between -7 and -222 for V18 and -70 and -368 for V14. The sequences conferring protein synthesis sensitivity were shown to reside between -502 and -61 of the H4 promoter.

摘要

我们研究了属于简单真核生物盘基网柄菌两类营养特异性基因的几个基因的表达和结构。在以细菌为食生长的变形虫中,所有营养特异性基因在发育开始时失活,到8至10小时时几乎不存在mRNA。相比之下,当细胞在无细菌肉汤中生长时,mRNA水平保持恒定,直到在10至12小时左右出现急剧下降。因此,在发育的最初几个小时内,这两类基因的调控都取决于先前的生长条件。对基因组克隆的分析已鉴定出两个V基因,V1和V18,为核糖体蛋白基因。未发现其他几个V基因是核糖体蛋白基因,这表明在盘基网柄菌中,非核糖体蛋白基因可能与核糖体蛋白基因协同调控。最后,通过缺失分析我们表明,两个V基因的启动子由位于这些基因调控表达所涉及序列上游的组成型正调控元件组成,对于V18在前545 bp上游,对于V14在前850 bp上游。V18参与调控表达的区域定位在-7至-222之间,V14在-70至-368之间。赋予蛋白质合成敏感性的序列显示位于H4启动子的-502至-61之间。

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