Thomas D, Rothstein R, Rosenberg N, Surdin-Kerjan Y
Laboratoire d'Enzymologie du Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Mol Cell Biol. 1988 Dec;8(12):5132-9. doi: 10.1128/mcb.8.12.5132-5139.1988.
In Saccharomyces cerevisiae the SAM1 and SAM2 genes encode two distinct forms of S-adenosylmethionine (AdoMet) synthetase. In a previous study we cloned and sequenced the SAM1 gene (D. Thomas and Y. Surdin-Kerjan, J. Biol. Chem. 262:16704-16709, 1987). In this work, the SAM2 gene was isolated by functional complementation of a yeast double-mutant strain, and its identity was ascertained by gene disruption. It has been sequenced and compared with the SAM1 gene. The degree of homology found between the two genes shows that SAM1 and SAM2 are duplicated genes. Using strains disrupted in one or the other SAM gene, we have studied the regulation of their expression by measuring the steady-state level of mRNA after growth under different conditions. The results show that the expression of the two SAM genes is regulated differently, SAM2 being induced by the presence of excess methionine in the growth medium and SAM1 being repressed under the same conditions. The level of mRNA in the parental strain shows that it is not the sum of the levels found in the two disrupted strains. This raises the question of how the two AdoMet synthetases in S. cerevisiae interact to control AdoMet synthesis.
在酿酒酵母中,SAM1和SAM2基因编码两种不同形式的S-腺苷甲硫氨酸(AdoMet)合成酶。在先前的一项研究中,我们克隆并测序了SAM1基因(D. 托马斯和Y. 苏尔丹-凯尔让,《生物化学杂志》262:16704 - 16709,1987年)。在这项工作中,通过对酵母双突变株进行功能互补分离出了SAM2基因,并通过基因破坏确定了其身份。已对其进行测序并与SAM1基因进行了比较。在这两个基因之间发现的同源程度表明SAM1和SAM2是重复基因。使用在其中一个或另一个SAM基因中被破坏的菌株,我们通过测量在不同条件下生长后mRNA的稳态水平来研究它们表达的调控。结果表明,这两个SAM基因的表达受到不同的调控,SAM2在生长培养基中存在过量甲硫氨酸时被诱导,而SAM1在相同条件下被抑制。亲代菌株中的mRNA水平表明它不是在两个被破坏菌株中发现的水平之和。这就提出了一个问题,即酿酒酵母中的两种AdoMet合成酶如何相互作用来控制AdoMet的合成。