Curcio M J, Sanders N J, Garfinkel D J
Bionetics Research, Inc., National Cancer Institute, Frederick, Maryland 21701.
Mol Cell Biol. 1988 Sep;8(9):3571-81. doi: 10.1128/mcb.8.9.3571-3581.1988.
Transposition of Ty elements in the yeast Saccharomyces cerevisiae occurs through an RNA intermediate. Although Ty RNA accounts for 5 to 10% of the total polyadenylated RNA in a haploid cell, the transposition frequency is only 10(-7) to 10(-8) per gene. To determine whether Ty elements native to the yeast genome are transpositionally competent, two elements were fused to the GAL1 promoter and tested for their ability to transpose. These native elements, Ty1-588 and Ty2-117, transposed at high levels when the GAL1 promoter was induced. Three Ty's identified as spontaneous transpositions in specific target genes were also tested. Of these three, Ty2-917 and the previously characterized element Ty1-H3 were shown to be transpositionally competent. The third element, Ty1-H1, was transposition defective. In addition, we marked the chromosomal copy of Ty1-588 with the NEO gene and demonstrated that Ty1-588NEO was actively transcribed in yeast cells. Ty1-588NEO transcription was regulated by the SPT3 and MAT loci in the same manner as that observed for Ty's collectively. These results indicate that the yeast genome contains functional Ty elements. The presence of a transpositionally competent, actively transcribed element suggests that regulation of Ty transposition occurs at a posttranscriptional level.
酵母酿酒酵母中Ty元件的转座通过RNA中间体发生。虽然Ty RNA在单倍体细胞中占总多聚腺苷酸化RNA的5%至10%,但每个基因的转座频率仅为10^(-7)至10^(-8)。为了确定酵母基因组中的天然Ty元件是否具有转座能力,将两个元件与GAL1启动子融合,并测试它们的转座能力。当GAL1启动子被诱导时,这些天然元件Ty1-588和Ty2-117会高水平转座。还测试了在特定靶基因中被鉴定为自发转座的三个Ty元件。在这三个元件中,Ty2-917和先前表征的元件Ty1-H3被证明具有转座能力。第三个元件Ty1-H1转座有缺陷。此外,我们用NEO基因标记了Ty1-588的染色体拷贝,并证明Ty1-588NEO在酵母细胞中被积极转录。Ty1-588NEO的转录受SPT3和MAT位点调控,调控方式与对Ty元件整体观察到的相同。这些结果表明酵母基因组包含功能性Ty元件。存在一个具有转座能力、被积极转录的元件表明Ty转座的调控发生在转录后水平。