Provost N M, Somers D E, Hurley J B
Department of Biochemistry, University of Washington, Seattle 98195.
J Biol Chem. 1988 Aug 25;263(24):12070-6.
A G protein alpha subunit gene has been isolated from a Drosophila melanogaster genomic library using a combination of bovine rod and cone transducin alpha subunit cDNAs as a probe under reduced stringency conditions. The gene, DG alpha 1, encodes a protein with an amino acid sequence 78% identical to bovine Gi alpha 1. However, unlike all reported Gi alpha subunit the DG alpha 1-encoded protein is not expected to be a pertussis toxin substrate, because it lacks a cysteine at the appropriate site. The protein coding region of the gene is split by four introns. The sequence of a head tissue cDNA clone, as well as amino acid similarities to mammalian G proteins, confirms this exon/intron structure. Northern blots of total cellular RNA reveal a major 2.3-kilobase transcript and a less abundant 1.7-kilobase transcript. These transcripts are most abundant in RNA from embryos and pupae. The DG alpha 1 gene is located on band 65C on the left arm of the third chromosome, on the basis of in situ hybridizations to Drosophila salivary gland polytene chromosomes.
在较低严谨性条件下,以牛视杆和视锥转导蛋白α亚基cDNA的混合物作为探针,从黑腹果蝇基因组文库中分离出一个G蛋白α亚基基因。该基因DGα1编码一种蛋白质,其氨基酸序列与牛Giα1有78%的同一性。然而,与所有已报道的Giα亚基不同,DGα1编码的蛋白质预计不是百日咳毒素的底物,因为它在适当位置缺少一个半胱氨酸。该基因的蛋白质编码区被四个内含子隔开。头部组织cDNA克隆的序列以及与哺乳动物G蛋白的氨基酸相似性证实了这种外显子/内含子结构。总细胞RNA的Northern印迹显示有一个主要的2.3千碱基转录本和一个较少的1.7千碱基转录本。这些转录本在胚胎和蛹的RNA中最为丰富。根据对果蝇唾液腺多线染色体的原位杂交,DGα1基因位于第三条染色体左臂的65C带。