Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico.
Mol Biol Cell. 2010 Feb 1;21(3):489-98. doi: 10.1091/mbc.e09-06-0472. Epub 2009 Dec 16.
The Kluyveromyces lactis heterotrimeric G protein is a canonical Galphabetagamma complex; however, in contrast to Saccharomyces cerevisiae, where the Ggamma subunit is essential for mating, disruption of the KlGgamma gene yielded cells with almost intact mating capacity. Expression of a nonfarnesylated Ggamma, which behaves as a dominant-negative in S. cerevisiae, did not affect mating in wild-type and DeltaGgamma cells of K. lactis. In contrast to the moderate sterility shown by the single DeltaKlGalpha, the double DeltaKlGalpha DeltaKlGgamma mutant displayed full sterility. A partial sterile phenotype of the DeltaKlGgamma mutant was obtained in conditions where the KlGbeta subunit interacted defectively with the Galpha subunit. The addition of a CCAAX motif to the C-end of KlGbeta, partially suppressed the lack of both KlGalpha and KlGgamma subunits. In cells lacking KlGgamma, the KlGbeta subunit cofractionated with KlGalpha in the plasma membrane, but in the DeltaKlGalpha DeltaKlGgamma strain was located in the cytosol. When the KlGbeta-KlGalpha interaction was affected in the DeltaKlGgamma mutant, most KlGbeta fractionated to the cytosol. In contrast to the generic model of G-protein function, the Gbeta subunit of K. lactis has the capacity to attach to the membrane and to activate mating effectors in absence of the Ggamma subunit.
乳酸克鲁维酵母三聚体 G 蛋白是一种典型的 Galpha-tagamma 复合物;然而,与酿酒酵母不同,酿酒酵母中的 Ggamma 亚基对于交配是必不可少的,而 KlGgamma 基因的破坏产生了几乎具有完整交配能力的细胞。表达一种非法呢基化的 Ggamma,它在酿酒酵母中表现为显性负性,不会影响野生型和 KlGgamma 基因缺失细胞的交配。与单 DeltaKlGalpha 表现出的中度不育相反,双 DeltaKlGalpha DeltaKlGgamma 突变体表现出完全不育。在 KlGbeta 亚基与 Galpha 亚基相互作用缺陷的条件下,DeltaKlGgamma 突变体获得了部分不育表型。KlGbeta 末端添加 CCAAX 基序部分抑制了 KlGalpha 和 KlGgamma 亚基的缺失。在缺乏 KlGgamma 的细胞中,KlGbeta 亚基与 KlGalpha 在质膜中共分馏,但在 DeltaKlGalpha DeltaKlGgamma 菌株中位于细胞质中。当 DeltaKlGgamma 突变体中 KlGbeta-KlGalpha 相互作用受到影响时,大多数 KlGbeta 分馏到细胞质中。与 G 蛋白功能的通用模型相反,在没有 Ggamma 亚基的情况下,Kl 乳杆菌的 Gbeta 亚基具有附着在膜上并激活交配效应物的能力。