Ke N, Irwin P A, Voytas D F
Department of Zoology and Genetics, Iowa State University, Ames, IA 50011, USA.
EMBO J. 1997 Oct 15;16(20):6272-80. doi: 10.1093/emboj/16.20.6272.
The Saccharomyces retrotransposon Ty5 integrates preferentially into transcriptionally inactive regions (silent chromatin) at the HM loci and telomeres. We found that silent chromatin represses basal Ty5 transcription, indicating that these elements are encompassed by silent chromatin in their native genomic context. Because transcription is a requirement for transposition, integration into silent chromatin would appear to prevent subsequent rounds of replication. Using plasmid-borne Ty5-lacZ constructs, we found that Ty5 expression is haploid specific and is repressed 10-fold in diploid strains. Ty5 transcription is also regulated by the pheromone response pathway and is induced approximately 20-fold upon pheromone treatment. Deletion analysis of the Ty5 LTR promoter revealed that a 33 bp region with three perfect matches to the pheromone response element is responsible for both mating pheromone and cell-type regulation. Transcriptional repression of Ty5 by silent chromatin can be reversed by pheromone treatment, which leads to transcription and transposition. Ty5 replication, therefore, is normally repressed by silent chromatin and appears to be induced during mating. This is the first example of transcriptional activation of a gene that naturally resides within silent chromatin.
酿酒酵母逆转座子Ty5优先整合到HM位点和端粒的转录非活性区域(沉默染色质)。我们发现沉默染色质抑制Ty5基础转录,这表明这些元件在其天然基因组环境中被沉默染色质所包围。由于转录是转座的必要条件,整合到沉默染色质中似乎会阻止后续的复制轮次。使用质粒携带的Ty5 - lacZ构建体,我们发现Ty5表达具有单倍体特异性,在二倍体菌株中被抑制10倍。Ty5转录也受信息素反应途径调控,在信息素处理后被诱导约20倍。对Ty5 LTR启动子的缺失分析表明,一个与信息素反应元件有三个完美匹配的33 bp区域负责交配信息素和细胞类型调控。沉默染色质对Ty5的转录抑制可通过信息素处理逆转,这会导致转录和转座。因此,Ty5复制通常被沉默染色质抑制,并且似乎在交配期间被诱导。这是自然存在于沉默染色质中的基因转录激活的第一个例子。