Bonaccorsi S, Gatti M, Pisano C, Lohe A
Centro di Genetica Evoluzionistica del CNR, Roma, Italy.
Chromosoma. 1990 Aug;99(4):260-6. doi: 10.1007/BF01731701.
Primary spermatocyte nuclei of Drosophila melanogaster exhibit three giant lampbrush-like loops formed by the kl-5, kl-3 and ks-1 Y chromosome fertility factors. Detailed mapping of satellite DNA sequences along the Y chromosome has recently shown that AA-GAC satellite repeats are a significant component of the kl-5 and ks-1 loop-forming regions. To determine whether these simple repeated sequences are transcribed on the loop structures we performed a series of DNA-RNA in situ hybridization experiments to fixed loop preparations using as a probe cloned AAGAC repeats. These experiments showed that the probe hybridizes with homologous transcripts specifically associated with the kl-5 and ks-1 loops. These transcripts are detected at all stages of development of these two loops, do not appear to migrate to the cytoplasm and are degraded when loops disintegrate during the first meiotic prophase. Moreover, an examination of the testes revealed that the transcription of the AAGAC sequences is restricted to the loops of primary spermatocytes; the other cell types of D. melanogaster spermatogenesis do not exhibit nuclear or cytoplasmic labeling. These experiments were confirmed by RNA blotting analysis which showed that transcription of the AAGAC sequences occurs in wild-type testes but not in X/O testes. The patterns of hybridization to the RNA blots indicated that the transcripts are highly heterogeneous in size, from large (migration at limiting mobility) to less than 1 kb. We discuss the possible function of the AAGAC satellite transcripts, in the light of the available information on the Y chromosome loops of D. melanogaster.
果蝇的初级精母细胞核呈现出由kl-5、kl-3和ks-1 Y染色体育性因子形成的三个巨大的灯刷样环。最近对Y染色体上卫星DNA序列的详细定位表明,AA-GAC卫星重复序列是kl-5和ks-1环形成区域的重要组成部分。为了确定这些简单重复序列是否在环结构上转录,我们使用克隆的AAGAC重复序列作为探针,对固定的环标本进行了一系列DNA-RNA原位杂交实验。这些实验表明,探针与特异性与kl-5和ks-1环相关的同源转录本杂交。在这两个环的所有发育阶段都能检测到这些转录本,它们似乎不会迁移到细胞质中,并且在第一次减数分裂前期环解体时会被降解。此外,对睾丸的检查表明,AAGAC序列的转录仅限于初级精母细胞的环;果蝇精子发生的其他细胞类型没有显示出核或细胞质标记。RNA印迹分析证实了这些实验,该分析表明AAGAC序列的转录发生在野生型睾丸中,而在X/O睾丸中不发生。与RNA印迹的杂交模式表明,转录本的大小高度异质,从大的(在极限迁移率下迁移)到小于1 kb。我们根据关于果蝇Y染色体环的现有信息讨论了AAGAC卫星转录本的可能功能。