Iwasaki M, Okumura K, Kondo Y, Tanaka T, Igarashi H
Shionogi Institute for Medical Science, Osaka, Japan.
Nucleic Acids Res. 1992 Aug 11;20(15):4001-7. doi: 10.1093/nar/20.15.4001.
The mammalian prion protein (PrPc) is a cellular protein of unknown function, an altered isoform of which (PrPsc) is a component of the infectious particle (prion) thought to be responsible for spongiform encephalopathies in humans and animals. The evolutionary conservation of the PrP gene has been reported in the genomes of many vertebrates as well as certain invertebrates. In the genome of nematode Caenorhabditis elegans, the sequence capable of hybridizing with the mammalian PrP cDNA probe has been demonstrated, predicting the presence of the PrP gene homologue in C.elegans. In this study, Southern analysis with the hamster PrP cDNA (HaPrP) probe confirmed the previous observation. Moreover, Northern analysis revealed that the sequence is actively transcribed in adult worms. Thus, we screened C.elegans cDNA libraries with the HaPrP probe and isolated a cDNA that hybridizes to the same sequence in C.elegans that hybridized with the HaPrP probe in the Southern and Northern analyses. The deduced amino acid sequence of this cDNA, however, is substantially homologous with heterogeneous nuclear ribonucleoprotein (hnRNP) core proteins rather than mammalian PrPc. The hnRNPs contain the glycine-rich domain in the C-terminal half of the molecule, which also seemed to be in PrPc at the N-terminal half of the molecule. Both of the glycine-rich domains are composed of tracts with high G + C content, indicating that these tracts may due to the hybridizing signals. These results suggest that this cDNA clone is derived from a novel hnRNP gene homologue in C.elegans but not from a predicted PrP gene homologue.
哺乳动物朊病毒蛋白(PrPc)是一种功能未知的细胞蛋白,其一种变异异构体(PrPsc)是感染性粒子(朊病毒)的组成成分,被认为是导致人类和动物海绵状脑病的原因。许多脊椎动物以及某些无脊椎动物的基因组中都报道了PrP基因的进化保守性。在线虫秀丽隐杆线虫的基因组中,已证明存在能够与哺乳动物PrP cDNA探针杂交的序列,这预示着秀丽隐杆线虫中存在PrP基因同源物。在本研究中,用仓鼠PrP cDNA(HaPrP)探针进行的Southern分析证实了先前的观察结果。此外,Northern分析表明该序列在成年线虫中活跃转录。因此,我们用HaPrP探针筛选了秀丽隐杆线虫cDNA文库,并分离出一个cDNA,它在秀丽隐杆线虫中与Southern和Northern分析中与HaPrP探针杂交的相同序列杂交。然而,该cDNA推导的氨基酸序列与异质性核核糖核蛋白(hnRNP)核心蛋白高度同源,而不是与哺乳动物PrPc同源。hnRNP在分子的C端一半含有富含甘氨酸的结构域,这在PrPc分子的N端一半似乎也存在。两个富含甘氨酸的结构域都由高G + C含量的片段组成,表明这些片段可能是杂交信号的原因。这些结果表明,这个cDNA克隆源自秀丽隐杆线虫中一个新的hnRNP基因同源物,而不是源自预测的PrP基因同源物。