Parisi M, Lin H
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Genetics. 1999 Sep;153(1):235-50. doi: 10.1093/genetics/153.1.235.
The pumilio (pum) gene plays an essential role in embryonic patterning and germline stem cell (GSC) maintenance during oogenesis in Drosophila. Here we report on a phenotypic analysis using pum(ovarette) mutations, which reveals multiple functions of pum in primordial germ cell proliferation, larval ovary formation, GSC division, and subsequent oogenic processes, as well as in oviposition. Specifically, by inducing pum(-) GSC clones at the onset of oogenesis, we show that pum is directly involved in GSC division, a function that is distinct from its requirement in primordial germ cells. Furthermore, we show that pum encodes 156- and 130-kD proteins, both of which are functional isoforms. Among pum(ovarette) mutations, pum(1688) specifically eliminates the 156-kD isoform but not the 130-kD isoform, while pum(2003) and pum(4277) specifically affect the 130-kD isoform but not the 156-kD isoform. Normal doses of both isoforms are required for the zygotic function of pum, yet either isoform alone at a normal dose is sufficient for the maternal effect function of pum. A pum cDNA transgene that contains the known open reading frame encodes only the 156-kD isoform and rescues the phenotype of both pum(1688) and pum(2003) mutants. These observations suggest that the 156- and 130-kD isoforms can compensate for each other's function in a dosage-dependent manner. Finally, we present molecular evidence suggesting that the two PUM isoforms share some of their primary structures.
在果蝇卵子发生过程中,pumilio(pum)基因在胚胎模式形成和生殖系干细胞(GSC)维持中起着至关重要的作用。在此,我们报告了一项使用pum(ovarette)突变的表型分析,该分析揭示了pum在原始生殖细胞增殖、幼虫卵巢形成、GSC分裂及随后的卵子发生过程以及产卵中的多种功能。具体而言,通过在卵子发生开始时诱导pum(-) GSC克隆,我们表明pum直接参与GSC分裂,这一功能与其在原始生殖细胞中的需求不同。此外,我们表明pum编码156-kD和130-kD蛋白,两者均为功能异构体。在pum(ovarette)突变中,pum(1688)特异性地消除了156-kD异构体但未消除130-kD异构体,而pum(2003)和pum(4277)特异性地影响130-kD异构体但未影响156-kD异构体。pum的合子功能需要两种异构体的正常剂量,但正常剂量的任何一种异构体单独就足以实现pum的母体效应功能。一个包含已知开放阅读框的pum cDNA转基因仅编码156-kD异构体,并挽救了pum(1688)和pum(2003)突变体的表型。这些观察结果表明,156-kD和130-kD异构体可以以剂量依赖的方式相互补偿功能。最后,我们提供了分子证据表明这两种PUM异构体共享一些一级结构。