Lochrie M A, Mendel J E, Sternberg P W, Simon M I
Division of Biology, California Institute of Technology, Pasadena 91125.
Cell Regul. 1991 Feb;2(2):135-54. doi: 10.1091/mbc.2.2.135.
A cDNA corresponding to a known G protein alpha subunit, the alpha subunit of Go (Go alpha), was isolated and sequenced. The predicted amino acid sequence of C. elegans Go alpha is 80-87% identical to other Go alpha sequences. An mRNA that hybridizes to the C. elegans Go alpha cDNA can be detected on Northern blots. A C. elegans protein that crossreacts with antibovine Go alpha antibody can be detected on immunoblots. A cosmid clone containing the C. elegans Go alpha gene (goa-1) was isolated and mapped to chromosome I. The genomic fragments of three other C. elegans G protein alpha subunit genes (gpa-1, gpa-2, and gpa-3) have been isolated using the polymerase chain reaction. The corresponding cosmid clones were isolated and mapped to disperse locations on chromosome V. The sequences of two of the genes, gpa-1 and gpa-3, were determined. The predicted amino acid sequences of gpa-1 and gpa-3 are only 48% identical to each other. Therefore, they are likely to have distinct functions. In addition they are not homologous enough to G protein alpha subunits in other organisms to be classified. Thus C. elegans has G proteins that are identifiable homologues of mammalian G proteins as well as G proteins that appear to be unique to C. elegans. Study of identifiable G proteins in C. elegans may result in a further understanding of their function in other organisms, whereas study of the novel G proteins may provide an understanding of unique aspects of nematode physiology.
分离并测序了与已知G蛋白α亚基(Go的α亚基,Goα)相对应的cDNA。秀丽隐杆线虫Goα的预测氨基酸序列与其他Goα序列的同一性为80 - 87%。在Northern印迹上可检测到与秀丽隐杆线虫Goα cDNA杂交的mRNA。在免疫印迹上可检测到与抗牛Goα抗体发生交叉反应的秀丽隐杆线虫蛋白。分离出了一个包含秀丽隐杆线虫Goα基因(goa - 1)的黏粒克隆,并将其定位到染色体I上。利用聚合酶链反应分离出了秀丽隐杆线虫其他三个G蛋白α亚基基因(gpa - 1、gpa - 2和gpa - 3)的基因组片段。分离出了相应的黏粒克隆,并将其定位到染色体V上的分散位置。测定了其中两个基因gpa - 1和gpa - 3的序列。gpa - 1和gpa - 3的预测氨基酸序列彼此间的同一性仅为48%。因此,它们可能具有不同的功能。此外,它们与其他生物中的G蛋白α亚基的同源性不足,无法进行分类。因此,秀丽隐杆线虫既有可鉴定为哺乳动物G蛋白同源物的G蛋白,也有似乎是秀丽隐杆线虫特有的G蛋白。对秀丽隐杆线虫中可鉴定的G蛋白的研究可能会进一步了解它们在其他生物中的功能,而对新型G蛋白的研究可能会有助于了解线虫生理学的独特方面。