Wu L, Valkema R, Van Haastert P J, Devreotes P N
Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205, USA.
J Cell Biol. 1995 Jun;129(6):1667-75. doi: 10.1083/jcb.129.6.1667.
Increasing evidence suggests that the beta gamma-subunit dimers of heterotrimeric G proteins play a pivotal role in transducing extracellular signals. The recent construction of G beta null mutants (g beta-) in Dictyostelium provides a unique opportunity to study the role of beta gamma dimers in signaling processes mediated by chemoattractant receptors. We have shown previously that g beta- cells fail to aggregate; in this study, we report the detailed characterization of these cells. The g beta- cells display normal motility but do not move towards chemattractants. The typical GTP-regulated high affinity chemoattractant-binding sites are lost in g beta- cells and membranes. The g beta- cells do not display chemoattractant-stimulated adenylyl cyclase or guanylyl cyclase activity. These results show that in vivo G beta links chemoattractant receptors to effectors and is therefore essential in many chemoattractant-mediated processes. In addition, we find that G beta is required for GTP gamma S stimulation of adenylyl cyclase activity, suggesting that the beta gamma-dimer activates the enzyme directly. Interestingly, the g beta- cells grow at the same rate as wild-type cells in axenic medium but grow more slowly on bacterial lawns and, therefore, may be defective in phagocytosis.
越来越多的证据表明,异源三聚体G蛋白的βγ亚基二聚体在转导细胞外信号中起关键作用。最近在盘基网柄菌中构建的Gβ缺失突变体(gβ-)为研究βγ二聚体在趋化因子受体介导的信号传导过程中的作用提供了独特的机会。我们之前已经表明,gβ-细胞无法聚集;在本研究中,我们报告了这些细胞的详细特征。gβ-细胞表现出正常的运动能力,但不会向趋化因子移动。典型的GTP调节的高亲和力趋化因子结合位点在gβ-细胞和膜中丢失。gβ-细胞不表现出趋化因子刺激的腺苷酸环化酶或鸟苷酸环化酶活性。这些结果表明,在体内Gβ将趋化因子受体与效应器联系起来,因此在许多趋化因子介导的过程中至关重要。此外,我们发现Gβ是GTPγS刺激腺苷酸环化酶活性所必需的,这表明βγ二聚体直接激活该酶。有趣的是,gβ-细胞在无菌培养基中的生长速度与野生型细胞相同,但在细菌草坪上生长较慢,因此可能在吞噬作用方面存在缺陷。