Chase Daniel L, Koelle Michael R
Department of Molecular Biophysics, The University of North Carolina at Chapel Hill, 27599-7260, USA.
Methods Enzymol. 2004;389:305-20. doi: 10.1016/S0076-6879(04)89018-9.
Caenorhabditis elegans has close homologs or orthologs of most mammalian (RGS) and G proteins, and mutants for all the RGS and G-protein genes of C. elegans have been generated. C. elegans RGS proteins can be matched to the specific Galpha proteins they regulate in vivo by comparing the defects in animals lacking or transgenically overexpressing an RGS protein with defects in a specific Galpha mutant. Transgenic expression of mutated RGS proteins or subdomains in C. elegans has also been used to carry out structure/function studies of RGS proteins. We propose that similar strategies can be used to understand the function of RGS proteins from other organisms by expressing them in C. elegans. This article describes general considerations regarding such experiments and provides detailed protocols for quantitatively measuring G-protein signaling phenotypes in C. elegans.
秀丽隐杆线虫拥有大多数哺乳动物（RGS）和G蛋白的紧密同源物或直系同源物，并且已经产生了秀丽隐杆线虫所有RGS和G蛋白基因的突变体。通过比较缺乏或转基因过表达RGS蛋白的动物中的缺陷与特定Gα突变体中的缺陷，可以将秀丽隐杆线虫RGS蛋白与其在体内调节的特定Gα蛋白相匹配。在秀丽隐杆线虫中突变RGS蛋白或亚结构域的转基因表达也已用于进行RGS蛋白的结构/功能研究。我们建议可以使用类似的策略，通过在秀丽隐杆线虫中表达来自其他生物体的RGS蛋白来了解其功能。本文描述了有关此类实验的一般注意事项，并提供了定量测量秀丽隐杆线虫中G蛋白信号表型的详细方案。
