Xu H, Boeke J D
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Mol Cell Biol. 1991 May;11(5):2736-43. doi: 10.1128/mcb.11.5.2736-2743.1991.
The Ty1 elements in the yeast Saccharomyces cerevisiae are a family of retrotransposons which transpose via a process similar to that of retroviral replication. We report here that the Ty1 transposition process can be blocked posttranscriptionally by treatment of cells with mating pheromones. When haploid yeast cells are treated with appropriate mating pheromones, the transposition frequency of a marked Ty1 element driven by the GAL1 promoter is greatly diminished. Ty1 viruslike particles (VLPs), the putative intermediates for transposition, can be isolated from mating pheromone-treated cells. These VLPs accumulate to normal levels but are aberrant in that they produce very few reverse transcripts of Ty1 RNA both in vivo and in vitro and contain subnormal amounts of p90-TYB and related proteins. In addition, a TYA phosphoprotein product accumulates in treated cells, and some species of TYB proteins have decreased stability. We also show that decreased transposition in mating pheromone-treated cells is not a consequence of simply blocking cell division, since Ty1 transposes at a nearly normal rate in yeast cells arrested in G2 by the drug nocodazole.
酿酒酵母中的Ty1元件是一类逆转座子,其转座过程类似于逆转录病毒的复制过程。我们在此报告,通过用交配信息素处理细胞,Ty1转座过程可在转录后被阻断。当单倍体酵母细胞用适当的交配信息素处理时,由GAL1启动子驱动的标记Ty1元件的转座频率会大大降低。Ty1病毒样颗粒(VLP)是转座的假定中间体,可从交配信息素处理的细胞中分离出来。这些VLP积累到正常水平,但存在异常,因为它们在体内和体外产生的Ty1 RNA逆转录产物很少,并且所含的p90-TYB及相关蛋白量低于正常水平。此外,一种TYA磷酸化蛋白产物在处理过的细胞中积累,并且某些种类的TYB蛋白稳定性降低。我们还表明,交配信息素处理过的细胞中转座减少并非简单地由于阻断细胞分裂所致,因为在被药物诺考达唑阻滞在G2期的酵母细胞中,Ty1以接近正常的速率转座。