Delidakis C, Kafatos F C
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
EMBO J. 1989 Mar;8(3):891-901. doi: 10.1002/j.1460-2075.1989.tb03450.x.
Drosophila melanogaster follicle cells over-replicate the chromosomal domain containing the third chromosome chorion gene cluster. Multiple regions of this cluster are needed in cis for attainment of high levels of amplification. We have confirmed the importance of the proposed amplification control element (ACE3) and demonstrated that it can support low levels of follicular amplification in the absence of other elements, but that it lacks detectable activity as a DNA replication origin. We have also demonstrated the existence of additional amplification-enhancing regions (AERs), by analyzing the amplification levels of a series of in situ induced, nested deletions of the chorion cluster. These deletions were induced by P-transposase perturbation of a chorion transposon in a highly amplifying transformed line, and were not accompanied by re-transposition, making possible a quantitative analysis of amplification levels in the absence of chromosomal position effects. Analysis of endogenous replication intermediates in wild-type follicular DNA suggested that at least one of the AERs may be an origin of replication and that amplification uses at least one additional replication origin.
黑腹果蝇的卵泡细胞会过度复制包含三号染色体绒毛膜基因簇的染色体区域。该基因簇的多个区域在顺式作用下对于实现高水平扩增是必需的。我们已经证实了所提出的扩增控制元件(ACE3)的重要性,并证明它在没有其他元件的情况下能够支持低水平的卵泡扩增,但它缺乏作为DNA复制起点的可检测活性。通过分析一系列原位诱导的绒毛膜基因簇嵌套缺失的扩增水平,我们还证明了存在额外的扩增增强区域(AER)。这些缺失是由高度扩增的转化系中绒毛膜转座子的P-转座酶扰动诱导的,并且没有伴随重新转座,从而使得在没有染色体位置效应的情况下对扩增水平进行定量分析成为可能。对野生型卵泡DNA中内源性复制中间体的分析表明,至少有一个AER可能是复制起点,并且扩增使用至少一个额外的复制起点。