Eki Toshihiko, Ishihara Takeshi, Katsura Isao, Hanaoka Fumio
Division of Life Science and Biotechnology, Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
DNA Res. 2007 Aug 31;14(4):183-99. doi: 10.1093/dnares/dsm016. Epub 2007 Oct 6.
Helicase-like proteins play a crucial role in nucleic acid- and chromatin-mediated reactions. In this study, we identified 134 helicase-like proteins in the nematode Caenorhabditis elegans and classified the proteins into 10 known subfamilies and a group of orphan genes on the basis of sequence similarity. We characterized loss-of-function phenotypes in RNA interference (RNAi)-treated animals for helicase family members, using the RNAi feeding method, and found several previously unreported phenotypes. Fifty-one (39.5%) of 129 genes tested showed development- or growth-defect phenotypes, and many of these genes were putative nematode homologs of essential genes in a unicellular eukaryote, budding yeast, suggesting conservation of these essential proteins in both species. Comparative analyses between these species identified evolutionarily diverged nematode proteins as well as conserved family members. Chromosome mapping of the nematode genes revealed 10 pairs of putative duplicated genes and clusters of C. elegans-specific SNF2-like genes and Helitrons. Analyses of transcriptional profile data revealed a predominantly oogenesis- and germline-enriched expression of many helicase-like genes. Finally, we identified the D2005.5(drh-3) gene in an RNAi-based screen for genes involved in resistance to X-ray irradiation. Analysis of DRH-3 will clarify the potentially novel mechanism by which it protects against X-ray-induced damage in C. elegans.
解旋酶样蛋白在核酸和染色质介导的反应中起着至关重要的作用。在本研究中,我们在秀丽隐杆线虫中鉴定出134种解旋酶样蛋白,并根据序列相似性将这些蛋白分为10个已知亚家族和一组孤儿基因。我们使用RNAi喂食法,对RNA干扰(RNAi)处理的动物中的解旋酶家族成员进行了功能缺失表型分析,发现了几种以前未报道的表型。在测试的129个基因中,有51个(39.5%)表现出发育或生长缺陷表型,其中许多基因是单细胞真核生物酿酒酵母中必需基因的假定线虫同源物,这表明这两种物种中这些必需蛋白具有保守性。这些物种之间的比较分析确定了进化上分化的线虫蛋白以及保守的家族成员。线虫基因的染色体定位揭示了10对假定的重复基因以及秀丽隐杆线虫特异性SNF2样基因和转座子Helitrons的簇。转录谱数据分析显示,许多解旋酶样基因主要在卵子发生和生殖系中富集表达。最后,我们在基于RNAi的筛选中鉴定出D2005.5(drh-3)基因,该基因参与对X射线辐射的抗性。对DRH-3进行分析将阐明其保护秀丽隐杆线虫免受X射线诱导损伤的潜在新机制。