Gillespie D E, Berg C A
University of Washington, Department of Genetics, Seattle 98195-7360, USA.
Genes Dev. 1995 Oct 15;9(20):2495-508. doi: 10.1101/gad.9.20.2495.
The homeless (hls) gene of Drosophila is required for anteroposterior and dorsoventral axis formation during oogenesis. At a low frequency, females homozygous for mutations in hls generate early egg chambers in which the oocyte is positioned incorrectly within the cyst. At a high frequency, late-stage egg chambers exhibit a ventralized chorion. Sequence analysis of the hls cDNA predicts a protein with amino-terminal homology to members of the DE-H family of RNA-dependent ATPases and putative helicases. Similarity of 51% in the amino-terminal third of the protein was found to two yeast splicing factors, PRP2 and PRP16, and to Drosophila Maleless, which is required for dosage compensation. To analyze Hls function, RNA localization patterns were determined for seven different transcripts in hls mutant ovaries. Previtellogenic transport to the oocyte was unaffected for all transcripts examined. Transport and localization of bicoid and oskar messages during vitellogenic stages were strongly disrupted, and the distribution and/or quantity of gurken, orb, and fs(1)K10 mRNAs were also affected, but to a lesser degree. In contrast, hu-li tai shao and Bicaudal-D transcripts were transported and localized normally in hls mutants. In addition, Kinesin heavy chain:beta-Galactosidase fusion protein failed to localize correctly to the posterior of the oocyte in vitellogenic egg chambers. Examination of the microtubule structure with anti-alpha-Tubulin antibodies revealed aberrant microtubule organizing center movement and an abnormally dense cytoplasmic microtubule meshwork. We discuss potential roles for Hls in organizing a cytoskeletal framework essential for localizing specific RNAs.
果蝇的无家可归(hls)基因在卵子发生过程中对前后轴和背腹轴的形成是必需的。在低频率下,hls突变纯合的雌性产生早期卵室,其中卵母细胞在囊肿内的定位不正确。在高频率下,晚期卵室表现出绒毛膜腹化。hls cDNA的序列分析预测一种蛋白质,其氨基末端与RNA依赖性ATP酶和推定解旋酶的DE-H家族成员具有同源性。在该蛋白质氨基末端三分之一处发现与两种酵母剪接因子PRP2和PRP16以及果蝇无雄性蛋白有51%的相似性,果蝇无雄性蛋白是剂量补偿所必需的。为了分析Hls的功能,确定了hls突变卵巢中七种不同转录本的RNA定位模式。对于所有检测的转录本,卵黄发生前向卵母细胞的运输不受影响。卵黄发生阶段期间,双尾和 Oskar 信息的运输和定位受到强烈破坏,并且 gurken、orb 和 fs(1)K10 mRNA 的分布和/或数量也受到影响,但程度较小。相比之下,hu-li tai shao和双尾-D转录本在hls突变体中运输和定位正常。此外,在卵黄发生的卵室中,驱动蛋白重链:β-半乳糖苷酶融合蛋白未能正确定位于卵母细胞的后部。用抗α-微管蛋白抗体检查微管结构,发现微管组织中心运动异常和细胞质微管网络异常密集。我们讨论了Hls在组织对特定RNA定位至关重要的细胞骨架框架中的潜在作用。