Lilly P, Wu L, Welker D L, Devreotes P N
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185.
Genes Dev. 1993 Jun;7(6):986-95. doi: 10.1101/gad.7.6.986.
Recent studies have demonstrated that G-protein-linked signal transduction pathways play a significant role in the developmental program of the simple eukaryotic organism Dictyostelium. We have reported previously the isolation of a G-protein beta-subunit and present here a more complete analysis of this gene. Low-stringency Southern blots and RFLP mapping studies suggest that the beta-subunit is a unique gene found on linkage group II. Its deduced amino acid sequence of 347 residues is approximately 60% identical to those of the human, Drosophila, and Caenorhabditis elegans beta-subunits. The carboxy-terminal 300 residues are about 70% identical; the amino-terminal 50 residues are quite divergent, containing only 10 identities. At all stages of growth and development, a single 1.9-kb beta-subunit mRNA is present at a high level, and a specific antibody detects a single 37-kD protein. We propose that G-protein heterotrimers are formed when this beta-subunit couples with each of the eight distinct G-protein alpha-subunits that are transiently expressed during development. Targeted disruption of the beta-subunit gene had no effect on the viability of haploid cells, but resulted in the inability of cells to aggregate.
最近的研究表明,G蛋白偶联信号转导途径在简单真核生物盘基网柄菌的发育程序中发挥着重要作用。我们之前报道过一种G蛋白β亚基的分离,在此展示对该基因更全面的分析。低严谨度的Southern印迹和RFLP图谱研究表明,β亚基是在连锁群II上发现的一个独特基因。其推导的347个残基的氨基酸序列与人类、果蝇和秀丽隐杆线虫的β亚基约60%相同。羧基末端的300个残基约70%相同;氨基末端的50个残基差异较大,仅含10个相同残基。在生长和发育的所有阶段,单一的1.9 kbβ亚基mRNA高水平存在,一种特异性抗体可检测到单一的37-kD蛋白。我们推测,当该β亚基与发育过程中瞬时表达的八个不同的G蛋白α亚基中的每一个偶联时,会形成G蛋白异源三聚体。β亚基基因的靶向破坏对单倍体细胞的活力没有影响,但导致细胞无法聚集。