G3 (Bethesda). 2012 Jun;2(6):657-63. doi: 10.1534/g3.112.002238. Epub 2012 Jun 1.
Suppressor screens are an invaluable method for identifying novel genetic interactions between genes in the model organism Caenorhabditis elegans. However, traditionally this approach has suffered from the laborious and protracted process of mapping mutations at the molecular level. Using a mutagen known to generate small deletions, coupled with oligoarray comparative genomic hybridization (aCGH), we have identified mutations in two genes that suppress the lethality associated with a mutation of the essential receptor tyrosine kinase rol-3. First, we find that deletion of the Bicaudal-C ortholog, bcc-1, suppresses rol-3-associated lethality. Second, we identify several duplications that also suppress rol-3-associated lethality. We establish that overexpression of srap-1, a single gene present in these duplications, mediates the suppression. This study demonstrates the suitability of deletion-biased mutagenesis screening in combination with aCGH characterization for the rapid identification of novel suppressor mutations. In addition to detecting small deletions, this approach is suitable for identifying copy number suppressor mutations, a class of suppressor not easily characterized using alternative approaches.
抑制筛选是鉴定秀丽隐杆线虫模型生物中基因之间新的遗传相互作用的一种非常有价值的方法。然而,传统上这种方法受到在分子水平上定位突变的繁琐和漫长过程的困扰。我们使用已知会产生小缺失的诱变剂,结合寡核苷酸阵列比较基因组杂交(aCGH),鉴定了两个基因的突变,这些突变可以抑制必需受体酪氨酸激酶 rol-3 突变相关的致死性。首先,我们发现 Bicaudal-C 同源物 bcc-1 的缺失可以抑制 rol-3 相关的致死性。其次,我们鉴定了几个也可以抑制 rol-3 相关致死性的重复。我们确定了这些重复中的一个基因 srap-1 的过表达介导了抑制作用。这项研究表明,在结合 aCGH 特征的情况下,偏向缺失的诱变筛选适合于快速鉴定新的抑制突变。除了检测小缺失外,这种方法还适合于鉴定拷贝数抑制突变,这是一类用替代方法不易表征的抑制突变。