Josse T, Boivin A, Anxolabéhère D, Ronsseray S
Laboratoire Dynamique du Génome et Evolution, Institut Jacques Monod, UMR7592, CNRS-Universités Paris 6 et 7, 2 place Jussieu, 75251 Paris Cedex 05, France.
Mol Genet Genomics. 2002 Nov;268(3):311-20. doi: 10.1007/s00438-002-0753-7. Epub 2002 Sep 25.
In Drosophila melanogaster, some clusters of P transgenes ( P-lacZ-white) display a variegating phenotype for the white marker in the eye, a phenomenon termed "Repeat-Induced Gene Silencing" (RIGS). We have tested the influence of the P element repression state (P cytotype) on the eye phenotype of several P-lac-w clusters that differ in transgene copy number or genomic insertion site. P element-encoded regulatory products strongly enhance RIGS. The effect occurs in both sexes, is detectable with clusters having at least three copies and is observed at both genomic locations tested (cytogenetic regions 50C and 92E). Single variegating P-lac-w transgenes located in pericentromeric heterochromatin are not affected by P regulatory products. All P strain backgrounds tested enhance RIGS, including chromosomes bearing a single P element encoding a truncated P transposase or carrying a single internally deleted KP element. Therefore, clusters are highly sensitive to different types of P repressors. Finally, a chimeric gene in which the 5' portion of the P element is fused to the polyhomeotic coding sequence (ph(p1)) also strongly enhances silencing of P-lac-w clusters. These results have implications for the mechanism of action of the P repressors and show that P transgene clusters represent a new class of P-sensitive alleles, providing a simple assay for somatic P repression that can be completed in one generation.
在黑腹果蝇中,一些P转座子基因簇(P-lacZ-白色)在眼睛中对于白色标记呈现出斑驳表型,这种现象被称为“重复诱导基因沉默”(RIGS)。我们测试了P元件抑制状态(P细胞型)对几个P-lac-w基因簇眼睛表型的影响,这些基因簇在转基因拷贝数或基因组插入位点上存在差异。P元件编码的调控产物强烈增强RIGS。这种效应在雌雄两性中均会出现,在具有至少三个拷贝的基因簇中可检测到,并且在测试的两个基因组位置(细胞遗传学区域50C和92E)均能观察到。位于着丝粒周围异染色质中的单个斑驳P-lac-w转基因不受P调控产物的影响。测试的所有P品系背景均增强RIGS,包括携带单个编码截短P转座酶的P元件的染色体或携带单个内部缺失的KP元件的染色体。因此,基因簇对不同类型的P抑制因子高度敏感。最后,一个嵌合基因,其中P元件的5'部分与多同源异型基因编码序列(ph(p1))融合,也强烈增强了P-lac-w基因簇的沉默。这些结果对P抑制因子的作用机制具有启示意义,并表明P转基因基因簇代表了一类新的P敏感等位基因,为可在一代内完成的体细胞P抑制提供了一种简单的检测方法。