Karpen G H, Spradling A C
Howard Hughes Medical Institute Research Laboratories, Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210.
Genetics. 1992 Nov;132(3):737-53. doi: 10.1093/genetics/132.3.737.
We investigated whether single P element insertional mutagenesis could be used to analyze heterochromatin within the Drosophila minichromosome Dp1187. Forty-five insertions of the P[lacZ,rosy+] element onto Dp1187 (recovered among 7,825 transpositions) were highly clustered. None was recovered in centromeric heterochromatin, but 39 occurred about 40 kb from the distal telomere within a 4.7-kb hotspot containing tandem copies of a novel 1.8-kb repetitive DNA sequence. The DNA within and distal to this region lacked essential genes and displayed several other properties characteristic of heterochromatin. The rosy+ genes within the inserted transposons were inhibited by position-effect variegation, and the subtelomeric region was underrepresented in polytene salivary gland cells. These experiments demonstrated that P elements preferentially transpose into a small subset of heterochromatic sites, providing a versatile method for studying the structure and function of these chromosome regions. This approach revealed that a Drosophila chromosome contains a large region of subtelomeric heterochromatin with specific structural and genetic properties.
我们研究了单个P因子插入诱变是否可用于分析果蝇小染色体Dp1187中的异染色质。P[lacZ,rosy+]因子在Dp1187上的45次插入(在7825次转座中获得)高度聚集。着丝粒异染色质中未获得任何插入,但有39次插入发生在距远端端粒约40 kb处的一个4.7 kb热点区域内,该热点区域包含一个新的1.8 kb重复DNA序列的串联拷贝。该区域内及其远端的DNA缺乏必需基因,并表现出异染色质的其他几个特征。插入的转座子中的rosy+基因受到位置效应斑驳的抑制,并且在多线唾液腺细胞中,亚端粒区域的代表性不足。这些实验表明,P因子优先转座到异染色质位点的一个小子集中,为研究这些染色体区域的结构和功能提供了一种通用方法。这种方法揭示了果蝇染色体包含一个具有特定结构和遗传特性的大亚端粒异染色质区域。