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首页> 外文期刊>Cell Division >Genetic analysis of the spindle checkpoint genes san-1, mdf-2, bub-3 and the CENP-F homologues hcp-1 and hcp-2 in Caenorhabditis elegans
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Genetic analysis of the spindle checkpoint genes san-1, mdf-2, bub-3 and the CENP-F homologues hcp-1 and hcp-2 in Caenorhabditis elegans

机译：秀丽隐杆线虫梭形检查点基因san-1，mdf-2，bub-3以及CENP-F同源物hcp-1和hcp-2的遗传分析

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Background The spindle checkpoint delays the onset of anaphase until all sister chromatids are aligned properly at the metaphase plate. To investigate the role san-1, the MAD3 homologue, has in Caenorhabditis elegans embryos we used RNA interference (RNAi) to identify genes synthetic lethal with the viable san-1(ok1580) deletion mutant. Results The san-1(ok1580) animal has low penetrating phenotypes including an increased incidence of males, larvae arrest, slow growth, protruding vulva, and defects in vulva morphogenesis. We found that the viability of san-1(ok1580) embryos is significantly reduced when HCP-1 (CENP-F homologue), MDF-1 (MAD-1 homologue), MDF-2 (MAD-2 homologue) or BUB-3 (predicted BUB-3 homologue) are reduced by RNAi. Interestingly, the viability of san-1(ok1580) embryos is not significantly reduced when the paralog of HCP-1, HCP-2, is reduced. The phenotype of san-1(ok1580);hcp-1(RNAi) embryos includes embryonic and larval lethality, abnormal organ development, and an increase in abnormal chromosome segregation (aberrant mitotic nuclei, anaphase bridging). Several of the san-1(ok1580);hcp-1(RNAi) animals displayed abnormal kinetochore (detected by MPM-2) and microtubule structure. The survival of mdf-2(RNAi);hcp-1(RNAi) embryos but not bub-3(RNAi);hcp-1(RNAi) embryos was also compromised. Finally, we found that san-1(ok1580) and bub-3(RNAi), but not hcp-1(RNAi) embryos, were sensitive to anoxia, suggesting that like SAN-1, BUB-3 has a functional role as a spindle checkpoint protein. Conclusion Together, these data suggest that in the C. elegans embryo, HCP-1 interacts with a subset of the spindle checkpoint pathway. Furthermore, the fact that san-1(ok1580);hcp-1(RNAi) animals had a severe viability defect whereas in the san-1(ok1580);hcp-2(RNAi) and san-1(ok1580);hcp-2(ok1757) animals the viability defect was not as severe suggesting that hcp-1 and hcp-2 are not completely redundant.
机译：背景纺锤体检查点延迟后期的开始，直到所有姐妹染色单体在中期板正确对齐为止。为了研究san-1（MAD3同源物）在秀丽隐杆线虫胚胎中的作用，我们使用RNA干扰（RNAi）鉴定了与可行的san-1（ok1580）缺失突变体合成致死的基因。结果san-1（ok1580）动物的穿透型低，包括雄性发病率增加，幼虫停滞，生长缓慢，外阴突出以及外阴形态发生缺陷。我们发现当HCP-1（CENP-F同源物），MDF-1（MAD-1同源物），MDF-2（MAD-2同源物）或BUB-3时，san-1（ok1580）胚胎的生存力显着降低。 （预测的BUB-3同源物）被RNAi还原。有趣的是，当减少HCP-1的旁系同源物HCP-2时，san-1（ok1580）胚胎的生存能力并未显着降低。 san-1（ok1580）; hcp-1（RNAi）胚胎的表型包括胚胎和幼虫致死率，异常器官发育以及异常染色体分离增加（有丝分裂核异常，后期桥接）。 san-1（ok1580）; hcp-1（RNAi）的几只动物显示异常的动粒（通过MPM-2检测）和微管结构。 mdf-2（RNAi）; hcp-1（RNAi）胚胎的存活，但不损害bub-3（RNAi）; hcp-1（RNAi）胚胎的存活。最后，我们发现san-1（ok1580）和bub-3（RNAi）胚胎对缺氧敏感，但对hcp-1（RNAi）则不敏感，这表明与SAN-1一样，BUB-3具有作为缺氧的功能。纺锤体检查点蛋白。结论总之，这些数据表明，在秀丽隐杆线虫胚胎中，HCP-1与纺锤体检查点途径的一部分相互作用。此外，san-1（ok1580）; hcp-1（RNAi）动物存在严重的生存能力缺陷，而在san-1（ok1580）; hcp-2（RNAi）和san-1（ok1580）; hcp- 2（ok1757）动物的生存能力缺陷没有那么严重，表明hcp-1和hcp-2并非完全多余。

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