Curcio M J, Hedge A M, Boeke J D, Garfinkel D J
BRI-Basic Research Program, NCI-Frederick Cancer Research Facility, MD, 21701.
Mol Gen Genet. 1990 Jan;220(2):213-21. doi: 10.1007/BF00260484.
To learn more about the variety of Ty elements capable of activating gene expression, we characterized 206 spontaneous Ty transpositions that activate the promoterless gene his3 delta 4. Most of the Ty elements appear to be full-length, although a few deleted elements were recovered. Over 95% of the insertions belong to the Ty1 family, and the rest are Ty2 elements. The excessive number of Ty1 transpositions was unexpected because there are only 2-fold more Ty1 than Ty2 elements in the yeast strains used in the selection. However, there is 20-fold more Ty1 than Ty2 RNA present in these yeast strains. This difference in RNA level explains the greater number of Ty1 verses Ty2 transpositions at his3 delta 4, because Ty elements transpose through an RNA intermediate. A similar association between the Ty transcript level and transpositional activation of his3 delta 4 is obtained in cells expressing GAL1-promoted Ty2-H556 or Ty2-917 elements, but only if the element does not contain a marker. Genetically marked Ty2-H556NEO and -917NEO elements transpose into and activate his3 delta 4 with the same efficiency as the previously characterized Ty1-H3NEO element, but are underrepresented relative to the levels of TyNEO transcript. We also found that chromosomal Ty transcripts are even more abundant than previously estimated and comprise about 1% of total cellular RNA.
为了更深入了解能够激活基因表达的各种Ty元件,我们对206个激活无启动子基因his3 delta 4的自发Ty转座进行了表征。尽管回收了一些缺失元件,但大多数Ty元件似乎是全长的。超过95%的插入属于Ty1家族,其余的是Ty2元件。Ty1转座数量过多出乎意料,因为在选择中使用的酵母菌株中Ty1元件仅比Ty2元件多2倍。然而,这些酵母菌株中存在的Ty1 RNA比Ty2 RNA多20倍。RNA水平的这种差异解释了在his3 delta 4处Ty1与Ty2转座数量更多的原因,因为Ty元件通过RNA中间体进行转座。在表达GAL1启动的Ty2 - H556或Ty2 - 917元件的细胞中,也获得了Ty转录水平与his3 delta 4的转座激活之间的类似关联,但前提是该元件不包含标记。带有遗传标记的Ty2 - H556NEO和 - 917NEO元件以与先前表征的Ty1 - H3NEO元件相同的效率转座并激活his3 delta 4,但相对于TyNEO转录本水平而言代表性不足。我们还发现染色体Ty转录本比先前估计的更加丰富,约占细胞总RNA的1%。