Greger I H, Proudfoot N J
Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
EMBO J. 1998 Aug 17;17(16):4771-9. doi: 10.1093/emboj/17.16.4771.
We have investigated transcriptional interactions between the GAL10 and GAL7 genes of Saccharomyces cerevisiae. Both genes are part of the galactose (GAL) gene cluster which is transcriptionally activated to high levels in the presence of galactose. Since GAL7 is positioned downstream of GAL10 and both genes are expressed co-ordinately at high levels, the possibility that GAL10 transcription influences GAL7 was analysed. Using transcriptional run-on assays, we show that high levels of polymerase are found in the 600 bp GAL10-7 intergenic region that accumulate over the GAL7 promoter. Furthermore, GAL7 transcription is enhanced when the GAL10 upstream activating sequence (UASG) is deleted, indicating that interference between GAL10 and GAL7 is likely to occur in the chromosomal locus. Deletions in the GAL10 poly(A) signal result in complete inactivation of the GAL7 promoter and cause a dramatic increase in bi-cistronic GAL10-7 mRNA, predominantly utilizing the downstream, GAL7 poly(A) site. These data demonstrate a pivotal role for the GAL10 poly(A) site in allowing the simultaneous expression of GAL10 and GAL7. In effect, this RNA processing signal has a direct influence on both transcriptional termination and initiation.
我们研究了酿酒酵母GAL10和GAL7基因之间的转录相互作用。这两个基因都是半乳糖(GAL)基因簇的一部分,该基因簇在半乳糖存在的情况下会被转录激活至高水平。由于GAL7位于GAL10的下游,且两个基因在高水平上协同表达,因此分析了GAL10转录影响GAL7的可能性。通过转录延伸分析,我们发现在GAL10 - 7基因间区域的600 bp处有高水平的聚合酶,这些聚合酶在GAL7启动子上积累。此外,当GAL10上游激活序列(UASG)缺失时,GAL7转录增强,这表明GAL10和GAL7之间的干扰可能发生在染色体位点。GAL10聚腺苷酸化信号的缺失导致GAL7启动子完全失活,并导致双顺反子GAL10 - 7 mRNA显著增加,主要利用下游的GAL7聚腺苷酸化位点。这些数据证明了GAL10聚腺苷酸化位点在允许GAL10和GAL7同时表达中起关键作用。实际上,这种RNA加工信号对转录终止和起始都有直接影响。