Smith P A, Corces V G
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Genetics. 1995 Jan;139(1):215-28. doi: 10.1093/genetics/139.1.215.
The gypsy retrotransposon of Drosophila melanogaster causes mutations that show temporal and tissue-specific phenotypes. These mutant phenotypes can be reversed by mutations in su(Hw), a gene that also regulates the transcription of the gypsy element. Gypsy encodes a full-length 7.0-kb RNA that is expressed in the salivary gland precursors and fat body of the embryo, imaginal discs and fat body of larvae, and fat body and ovaries of adult females. The su(Hw)-binding region inserted upstream of the promoter of a lacZ reporter gene can induce beta-galactosidase expression in a subset of the embryonic and larval tissues where gypsy is normally transcribed. This expression is dependent on the presence of a functional su(Hw) product, suggesting that this protein is a positive activator of gypsy transcription. Flies transformed with a construct in which the 5' LTR and leader sequences of gypsy are fused to lacZ show beta-galactosidase expression in all tissues where gypsy is normally expressed, indicating that sequences other than the su(Hw)-binding site are required for proper spatial and temporal expression of gypsy. Mutations in the zinc fingers of su(Hw) affect its ability to bind DNA and to induce transcription of the lacZ reporter gene. Two other structural domains of su(Hw) also play an important role in transcriptional regulation of gypsy. Deletion of the amino-terminal acidic domain results in the loss of lacZ expression in larval fat body and adult ovaries, whereas mutations in the leucine zipper region result in an increase of lacZ expression in larval fat body and a decrease in adult ovaries. These effects might be the result of interactions of su(Hw) with activator and repressor proteins through the acidic and leucine zipper domains to produce the final pattern of tissue-specific expression of gypsy.
黑腹果蝇的吉普赛逆转座子会引发一些突变,这些突变呈现出时间和组织特异性的表型。这些突变表型可被su(Hw)基因的突变所逆转,su(Hw)基因也调控着吉普赛元件的转录。吉普赛编码一种全长7.0 kb的RNA,它在胚胎的唾液腺前体和脂肪体、幼虫的成虫盘和脂肪体以及成年雌性的脂肪体和卵巢中表达。插入到lacZ报告基因启动子上游的su(Hw)结合区域,可在吉普赛正常转录的部分胚胎和幼虫组织中诱导β-半乳糖苷酶表达。这种表达依赖于功能性su(Hw)产物的存在,这表明该蛋白是吉普赛转录的正激活因子。用一种构建体转化果蝇,其中吉普赛的5' LTR和前导序列与lacZ融合,结果显示在吉普赛正常表达的所有组织中都有β-半乳糖苷酶表达,这表明除了su(Hw)结合位点之外的序列对于吉普赛正确的时空表达是必需的。su(Hw)锌指结构域的突变会影响其结合DNA以及诱导lacZ报告基因转录的能力。su(Hw)的另外两个结构域在吉普赛的转录调控中也起着重要作用。氨基末端酸性结构域的缺失导致幼虫脂肪体和成年卵巢中lacZ表达丧失,而亮氨酸拉链区域的突变导致幼虫脂肪体中lacZ表达增加,成年卵巢中lacZ表达减少。这些效应可能是su(Hw)通过酸性和亮氨酸拉链结构域与激活蛋白和抑制蛋白相互作用,从而产生吉普赛组织特异性表达最终模式的结果。