Large-scale RNAi screens identify novel genes that interact with the C. elegans retinoblastoma pathway as well as splicing-related components with synMuv B activity.
Large-scale RNAi screens identify novel genes that interact with the C. elegans retinoblastoma pathway as well as splicing-related components with synMuv B activity.
复制标题
DOI：
10.1186/1471-213x-7-30
复制
发表时间：
2007-04-06
影响因子：
--
通讯作者：
van den Heuvel, Sander
中科院分区：
文献类型：
--
作者：
Ceron, Julian;Rual, Jean-Francois;Chandra, Abha;Dupuy, Denis;Vidal, Marc;van den Heuvel, Sander
视网膜母细胞瘤肿瘤抑制因子（Rb）在大多数人类癌症中不受调控的保守途径中起作用。秀丽隐杆线虫（Caenorhabditis elegans）中单个rb相关基因lin-35的失活对生存能力和生育能力的影响有限，但当与特定其他基因的失活结合时，会导致细胞命运和细胞周期调节的变化。例如，lin- 35rb是一个合成的多外阴（synMuv） B类基因，当与a类或C类synMuv基因同时失活时，会导致多外阴表型。我们使用ORFeome RNAi文库来鉴定与秀丽隐杆线虫lin-35 Rb相互作用的基因，并鉴定出与rrf-3和eri-1 RNAi敏感突变体相比，在lin-35突变体中显示合成或增强RNAi表型的57个基因。基于缺失等位基因的特征，发现合成的lin-35相互作用子zfp-2抑制RNAi，并与lin-35 Rb在性腺发育中协同作用。有趣的是，十个剪接相关基因的功能与lin- 35rb相似，作为防止不适当外阴诱导的synMuv B基因。特定剪接体组分的部分失活进一步揭示了lin- 35rb在细胞周期控制、转基因表达和种系颗粒限制性表达方面的功能相似性。我们鉴定了一系列候选的lin-35 Rb相互作用基因，并验证了zfp-2是一个新的lin-35合成致死基因。此外，我们观察到lin-35 rb控制过程中一个剪接组件子集的新作用。我们的数据支持关于抗癌治疗和基因表达多级调控可能性的新假设。
The retinoblastoma tumor suppressor (Rb) acts in a conserved pathway that is deregulated in most human cancers. Inactivation of the single Rb-related gene in Caenorhabditis elegans, lin-35, has only limited effects on viability and fertility, yet causes changes in cell-fate and cell-cycle regulation when combined with inactivation of specific other genes. For instance, lin-35 Rb is a synthetic multivulva (synMuv) class B gene, which causes a multivulva phenotype when inactivated simultaneously with a class A or C synMuv gene. We used the ORFeome RNAi library to identify genes that interact with C. elegans lin-35 Rb and identified 57 genes that showed synthetic or enhanced RNAi phenotypes in lin-35 mutants as compared to rrf-3 and eri-1 RNAi hypersensitive mutants. Based on characterizations of a deletion allele, the synthetic lin-35 interactor zfp-2 was found to suppress RNAi and to cooperate with lin-35 Rb in somatic gonad development. Interestingly, ten splicing-related genes were found to function similar to lin-35 Rb, as synMuv B genes that prevent inappropriate vulval induction. Partial inactivation of specific spliceosome components revealed further similarities with lin-35 Rb functions in cell-cycle control, transgene expression and restricted expression of germline granules. We identified an extensive series of candidate lin-35 Rb interacting genes and validated zfp-2 as a novel lin-35 synthetic lethal gene. In addition, we observed a novel role for a subset of splicing components in lin-35 Rb-controlled processes. Our data support novel hypotheses about possibilities for anti-cancer therapies and multilevel regulation of gene expression.