Xue Chaoyang, Hsueh Yen-Ping, Chen Lydia, Heitman Joseph
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
Mol Microbiol. 2008 Oct;70(2):379-95. doi: 10.1111/j.1365-2958.2008.06417.x. Epub 2008 Aug 27.
G proteins orchestrate critical cellular functions by transducing extracellular signals into internal signals and controlling cellular responses to environmental cues. G proteins typically function as switches that are activated by G protein-coupled receptors (GPCRs) and negatively controlled by regulator of G protein signalling (RGS) proteins. In the human fungal pathogen Cryptococcus neoformans, three G protein alpha subunits (Gpa1, Gpa2 and Gpa3) have been identified. In a previous study, we identified the RGS protein Crg2 involved in regulating the pheromone response pathway through Gpa2 and Gpa3. In this study, a role for Crg2 was established in the Gpa1-cAMP signalling pathway that governs mating and virulence. We show that Crg2 physically interacts with Gpa1 and crg2 mutations increase cAMP production. crg2 mutations also enhance mating filament hyphae production, but reduce cell-cell fusion and sporulation efficiency during mating. Although crg2 mutations and the Gpa1 dominant active allele GPA1(Q284L) enhanced melanin production under normally repressive conditions, virulence was attenuated in a murine model. We conclude that Crg2 participates in controlling both Gpa1-cAMP-virulence and pheromone-mating signalling cascades and hypothesize it may serve as a molecular interface between these two central signalling conduits.
G蛋白通过将细胞外信号转化为细胞内信号并控制细胞对环境线索的反应,来协调关键的细胞功能。G蛋白通常作为开关发挥作用,由G蛋白偶联受体(GPCRs)激活,并受到G蛋白信号调节剂(RGS)蛋白的负调控。在人类真菌病原体新型隐球菌中,已鉴定出三种G蛋白α亚基(Gpa1、Gpa2和Gpa3)。在先前的一项研究中,我们鉴定出RGS蛋白Crg2参与通过Gpa2和Gpa3调节信息素反应途径。在本研究中,确定了Crg2在控制交配和毒力的Gpa1-cAMP信号通路中的作用。我们发现Crg2与Gpa1发生物理相互作用,且crg2突变会增加cAMP的产生。crg2突变还会增强交配丝状菌丝的产生,但会降低交配过程中的细胞间融合和孢子形成效率。尽管crg2突变和Gpa1显性活性等位基因GPA1(Q284L)在正常抑制条件下会增强黑色素的产生,但在小鼠模型中毒力会减弱。我们得出结论,Crg2参与控制Gpa1-cAMP-毒力和信息素-交配信号级联反应,并推测它可能作为这两个中心信号传导途径之间的分子界面。