Rai R, Daugherty J R, Cunningham T S, Cooper T G
Department of Microbiology and Immunology, University of Tennessee, Memphis, Tennessee 38163, USA.
J Biol Chem. 1999 Sep 24;274(39):28026-34. doi: 10.1074/jbc.274.39.28026.
Allantoin pathway gene expression in Saccharomyces cerevisiae responds to two different environmental stimuli. The expression of these genes is induced in the presence of allantoin or its degradative metabolites and repressed when a good nitrogen source (e. g. asparagine or glutamine) is provided. Three types of cis-acting sites and trans-acting factors are required for allantoin pathway gene transcription as follows: (i) UAS(NTR) element associated with the transcriptional activators Gln3p and Gat1p, (ii) URS(GATA) element associated with the repressor Dal80p, and (iii) UIS(ALL) element associated with the Dal82 and Dal81 proteins required for inducer-dependent transcription. Most of the work leading to the above conclusions has employed inducer-independent allantoin pathway genes (e.g. DAL5 and DAL3). The purpose of this work is to extend our understanding of these elements and their roles to inducible allantoin pathway genes using the DAL7 (encoding malate synthase) as a model. We show that eight distinct cis-acting sites participate in the process as follows: a newly identified GC-rich element, two UAS(NTR), two UIS(ALL), and three URS(GATA) elements. The two GATA-containing UAS(NTR) elements are coincident with two of the three GATA sequences that make up the URS(GATA) elements. The remaining URS(GATA) GATA sequence, however, is not a UAS(NTR) element but appears to function only in repression. The data provide insights into how these cis- and trans-acting factors function together to accomplish the regulated expression of the DAL7 gene that is observed in vivo.
酿酒酵母中尿囊素途径基因的表达对两种不同的环境刺激做出反应。这些基因的表达在尿囊素或其降解代谢产物存在时被诱导,而当提供优质氮源(如天冬酰胺或谷氨酰胺)时则被抑制。尿囊素途径基因转录需要三种类型的顺式作用位点和反式作用因子,具体如下:(i)与转录激活因子Gln3p和Gat1p相关的UAS(NTR)元件;(ii)与阻遏蛋白Dal80p相关的URS(GATA)元件;(iii)与诱导物依赖性转录所需的Dal82和Dal81蛋白相关的UIS(ALL)元件。得出上述结论的大部分研究工作都使用了不依赖诱导物的尿囊素途径基因(如DAL5和DAL3)。本研究的目的是将我们对这些元件及其作用的理解扩展到以DAL7(编码苹果酸合酶)为模型的可诱导尿囊素途径基因。我们发现八个不同的顺式作用位点参与了这一过程,具体如下:一个新鉴定的富含GC的元件、两个UAS(NTR)、两个UIS(ALL)和三个URS(GATA)元件。两个含GATA的UAS(NTR)元件与构成URS(GATA)元件的三个GATA序列中的两个重合。然而,其余的URS(GATA) GATA序列不是UAS(NTR)元件,似乎仅在抑制中起作用。这些数据为这些顺式和反式作用因子如何共同发挥作用以实现体内观察到的DAL7基因的调控表达提供了见解。