Yu G, Fassler J S
Department of Biology, University of Iowa, Iowa City 52242.
Mol Cell Biol. 1993 Jan;13(1):63-71. doi: 10.1128/mcb.13.1.63-71.1993.
The Ty transposable elements of Saccharomyces cerevisiae consist of a single large transcription unit whose expression is controlled by a combination of upstream and downstream regulatory sequences. Errede (B. Errede, Mol. Cell. Biol. 13:57-62, 1993) has shown that among the downstream control sequences is a binding site for the transcription factor, MCM1. A small restriction fragment containing the Ty1 MCM1-binding site exhibits very weak activation of heterologous gene expression. The absence of SPT13 (GAL11) causes a dramatic increase in activity directed by these sequences. This effect is mediated through the MCM1-binding site itself. MCM1 mRNA and protein levels, as well as its affinity for its binding site, are unchanged in the absence of SPT13. Our results suggest that SPT13 has a role in the negative control of MCM1 activity that is likely to be posttranslational. A role for SPT13 in the negative regulation of the activity of the Ty1 MCM1-binding site is consistent with our previous proposal that spt13-mediated suppression of Ty insertion mutations could be attributed to the loss of negative regulation of genes adjacent to Ty elements.
酿酒酵母的Ty转座元件由单个大的转录单元组成,其表达受上游和下游调控序列组合的控制。埃雷德(B. 埃雷德,《分子与细胞生物学》13:57 - 62,1993年)已表明,在下游控制序列中有转录因子MCM1的结合位点。一个包含Ty1 MCM1结合位点的小限制性片段对异源基因表达的激活作用非常微弱。SPT13(GAL11)的缺失会导致这些序列所介导的活性急剧增加。这种效应是通过MCM1结合位点本身介导的。在没有SPT13的情况下,MCM1的mRNA和蛋白质水平及其与结合位点的亲和力均未改变。我们的结果表明,SPT13在MCM1活性的负调控中起作用,这可能是在翻译后水平。SPT13在Ty1 MCM1结合位点活性的负调控中的作用与我们之前的提议一致,即spt13介导的对Ty插入突变的抑制可能归因于与Ty元件相邻基因的负调控丧失。