Keating Christopher D, Kriek Neline, Daniels Margaret, Ashcroft Neville R, Hopper Neil A, Siney Elodie J, Holden-Dye Lindy, Burke Julian F
Department of Biochemistry, University of Sussex, Falmer, East Sussex BN1 9QG, United Kingdom.
Curr Biol. 2003 Sep 30;13(19):1715-20. doi: 10.1016/j.cub.2003.09.003.
G protein-coupled receptors (GPCRs) are the largest family of genes in animal genomes and represent more than 2% of genes in humans and C. elegans. These evolutionarily conserved seven-transmembrane proteins transduce a diverse range of signals. In view of their pivotal role in cell signaling, it is perhaps surprising that decades of genetic analysis in C. elegans, and recent genome-wide RNAi screens, have identified very few GPCR mutants. Therefore, we screened all GPCRs predicted to bind either small-molecule neurotransmitters or neuropeptides by using RNAi and quantitative behavioral assays. This shows that C16D6.2, C25G6.5, C26F1.6, F35G8.1, F41E7.3, and F59C12.2 are likely to be involved in reproduction, whereas C15B12.5, C10C6.2, C24A8.4, F15A8.5, F59D12.1, T02E9.1, and T05A1.1 have a role in locomotion. Gene deletions for F35G8.1 and T05A1.1 resulted in the same phenotype as that seen with RNAi. As some GPCRs may be resistant to RNAi, or may result in abnormalities not screened for here, the actual proportion of nonredundant receptors with an assayable function is probably greater. Strikingly, most phenotypes were observed for NPY-like receptors that may bind neuropeptides. This is consistent with the known actions of neuropeptides on the body wall muscle and reproductive tract in nematodes.
G蛋白偶联受体(GPCRs)是动物基因组中最大的基因家族,在人类和秀丽隐杆线虫中占基因总数的2%以上。这些在进化上保守的七跨膜蛋白可转导多种信号。鉴于它们在细胞信号传导中的关键作用,也许令人惊讶的是,在秀丽隐杆线虫中进行了数十年的遗传分析以及最近的全基因组RNA干扰筛选,却只鉴定出极少数的GPCR突变体。因此,我们通过RNA干扰和定量行为分析筛选了所有预测可结合小分子神经递质或神经肽的GPCR。结果表明,C16D6.2、C25G6.5、C26F1.6、F35G8.1、F41E7.3和F59C12.2可能参与生殖,而C15B12.5、C10C6.2、C24A8.4、F15A8.5、F59D12.1、T02E9.1和T05A1.1在运动中起作用。F35G8.1和T05A1.1的基因缺失导致了与RNA干扰相同的表型。由于一些GPCR可能对RNA干扰有抗性,或者可能导致此处未筛选到的异常情况,具有可检测功能的非冗余受体的实际比例可能更高。引人注目的是,大多数表型是在可能结合神经肽的NPY样受体中观察到的。这与神经肽对线虫体壁肌肉和生殖道的已知作用一致。