Ahn Sejin, Spatt Dan, Winston Fred
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Eukaryot Cell. 2012 Aug;11(8):1067-74. doi: 10.1128/EC.00141-12. Epub 2012 Jun 15.
The Schizosaccharomyces pombe inv1(+) gene encodes invertase, the enzyme required for hydrolysis of sucrose and raffinose. Transcription of inv1(+) is regulated by glucose levels, with transcription tightly repressed in high glucose and strongly induced in low glucose. To understand this regulation, we have analyzed the inv1(+) cis-regulatory region and the requirement for the trans-acting coactivators SAGA and Swi/Snf. Surprisingly, deletion of the entire 1-kilobase intergenic region between the inv1(+) TATA element and the upstream open reading frame SPCC191.10 does not significantly alter regulation of inv1(+) transcription. However, a longer deletion that extends through SPCC191.10 abolishes inv1(+) induction in low glucose. Additional analysis demonstrates that there are multiple, redundant regulatory regions spread over 1.5 kb 5' of inv1(+), including within SPCC191.10, that can confer glucose-mediated transcriptional regulation to inv1(+). Furthermore, SPCC191.10 can regulate inv1(+) transcription in an orientation-independent fashion and from a distance as great as 3 kb. With respect to trans-acting factors, both SAGA and Swi/Snf are recruited to SPCC191.10 and to other locations in the large inv1(+) regulatory region in a glucose-dependent fashion, and both are required for inv1(+) derepression. Taken together, these results demonstrate that inv1(+) regulation in S. pombe occurs via the use of multiple regulatory elements and that activation can occur over a great distance, even from elements within other open reading frames.
粟酒裂殖酵母inv1(+)基因编码转化酶,这是水解蔗糖和棉子糖所需的酶。inv1(+)的转录受葡萄糖水平调控,在高葡萄糖条件下转录被紧密抑制,在低葡萄糖条件下则被强烈诱导。为了解这种调控机制,我们分析了inv1(+)的顺式调控区域以及对反式作用共激活因子SAGA和Swi/Snf的需求。令人惊讶的是,删除inv1(+) TATA元件与上游开放阅读框SPCC191.10之间的整个1千碱基基因间区域,并不会显著改变inv1(+)转录的调控。然而,延伸至SPCC191.10的更长缺失会消除低葡萄糖条件下inv1(+)的诱导。进一步分析表明,在inv1(+)上游1.5 kb范围内分布着多个冗余调控区域,包括SPCC191.10内部,这些区域可赋予inv1(+)葡萄糖介导的转录调控。此外,SPCC191.10能够以方向独立的方式并在高达3 kb的距离外调控inv1(+)转录。关于反式作用因子,SAGA和Swi/Snf都以葡萄糖依赖的方式被招募到SPCC191.10以及大的inv1(+)调控区域中的其他位置,并且两者都是inv1(+)去抑制所必需的。综上所述,这些结果表明粟酒裂殖酵母中inv1(+)的调控是通过使用多个调控元件实现的,并且激活可以在很远的距离发生,甚至可以从其他开放阅读框内的元件发生。