Steller H, Pirrotta V
Mol Cell Biol. 1986 May;6(5):1640-9. doi: 10.1128/mcb.6.5.1640-1649.1986.
We have transformed Drosophila melanogaster with modified P-element transposons, which express the transposase function from the heat-inducible hsp70 heat shock promoter. The Icarus transposon, which contains a direct hsp70-P fusion gene, behaved like a very active autonomous P element even before heat shock induction. Although heat shock led to abundant somatic transcription, transposition of the Icarus element was confined to germ line cells. To reduce the constitutive transposase activity observed for the Icarus element, we attenuated the translational efficiency of the transposase RNA by inserting the transposon 5 neomycin resistance gene between the hsp70 promoter and the P-element sequences. The resulting construct, called Icarus-neo, conferred resistance to G418, and its transposition was significantly stimulated by heat shock. Heat shocks applied during the embryonic or third instar larval stage had similar effects, indicating that transposition of P elements is not restricted to a certain developmental stage. Both Icarus and Icarus-neo destabilized snw in a P-cytotype background and thus at least partially overcome the repression of transposition. Our results suggest that the regulation of P-element transposition occurs at both the transcriptional and posttranscriptional levels.
我们用修饰过的P因子转座子转化了黑腹果蝇,这些转座子从热诱导型hsp70热休克启动子表达转座酶功能。伊卡洛斯转座子含有一个直接的hsp70 - P融合基因,甚至在热休克诱导之前就表现得像一个非常活跃的自主P因子。尽管热休克导致了大量的体细胞转录,但伊卡洛斯元件的转座仅限于生殖系细胞。为了降低伊卡洛斯元件观察到的组成型转座酶活性,我们通过在hsp70启动子和P因子序列之间插入转座子5新霉素抗性基因来减弱转座酶RNA的翻译效率。得到的构建体称为伊卡洛斯 - neo,赋予了对G418的抗性,其转座受到热休克的显著刺激。在胚胎期或三龄幼虫期施加热休克有相似的效果,表明P因子的转座不限于某个发育阶段。在P细胞质型背景下,伊卡洛斯和伊卡洛斯 - neo都使snw不稳定,因此至少部分克服了转座的抑制。我们的结果表明,P因子转座的调控发生在转录和转录后水平。