Crabeel M, Lavalle R, Glansdorff N
Microbiology, Vrije Universiteit Brussel, Belgium.
Mol Cell Biol. 1990 Mar;10(3):1226-33. doi: 10.1128/mcb.10.3.1226-1233.1990.
A specific repression mechanism regulates arginine biosynthesis in Saccharomyces cerevisiae. The involvement of regulatory proteins displaying DNA-binding features and the location of an operator region between the TATA box and the transcription start of the structural gene ARG3 suggest that this mechanism operates at the level of transcription. A posttranscriptional mechanism has, however, been proposed to account for the conspicuous lack of proportionality between ARG3 mRNA steady-state levels (as determined by Northern [RNA] assays; F. Messenguy and E. Dubois, Mol. Gen. Genet. 189:148-156, 1983) and the cognate enzyme activities. In this work, we have analyzed the time course of the incorporation of radioactive precursors into ARG1 and ARG3 mRNAs and the kinetics of their decay under different regulatory statuses. The results (expressed in terms of relative mRNA levels, relative transcription rates, and mRNA half-lives) give the picture expected from a purely transcriptional control. A similar analysis of expression of the gene CPA1, for which a translational regulation by arginine has been clearly demonstrated (M. Werner, A. Feller, F. Messenguy, and A. Piérard, Cell 49:805-813, 1987), indicates that this gene is also partly regulated at the transcriptional level by the ARGR repressor system. Moreover, the half-life of CPA1 mRNA is reduced twofold in the presence of excess arginine; we suggest that this could be inherent in the mechanism of translational regulation of CPA1.
一种特定的抑制机制调控酿酒酵母中的精氨酸生物合成。具有DNA结合特性的调控蛋白的参与以及操纵区位于结构基因ARG3的TATA框和转录起始点之间,这表明该机制在转录水平起作用。然而,有人提出一种转录后机制来解释ARG3 mRNA稳态水平(通过Northern[RNA]分析确定;F. Messenguy和E. Dubois,《分子与普通遗传学》189:148 - 156,1983)与相应酶活性之间明显缺乏比例关系的现象。在这项工作中,我们分析了放射性前体掺入ARG1和ARG3 mRNA的时间进程以及它们在不同调控状态下的衰变动力学。结果(以相对mRNA水平、相对转录速率和mRNA半衰期表示)呈现出纯转录调控所预期的情况。对基因CPA1表达的类似分析表明,该基因也部分受ARGR阻遏系统在转录水平的调控,此前已明确证明精氨酸对其有翻译调控作用(M. Werner、A. Feller、F. Messenguy和A. Piérard,《细胞》49:805 - 813,1987)。此外,在精氨酸过量的情况下,CPA1 mRNA的半衰期缩短了两倍;我们认为这可能是CPA1翻译调控机制所固有的。