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从青霉属(Penicillium chrysogenum)中分离得到的异三聚体 Gα 蛋白 Pga1 响应碳源触发萌发,并对不同应激条件的抗性产生负面影响。

Heterotrimeric Gα protein Pga1 from Penicillium chrysogenum triggers germination in response to carbon sources and affects negatively resistance to different stress conditions.

机构信息

Institute of Biotechnology of León (INBIOTEC), Parque Científico de León, Av. Real 1, 24006 León, Spain.

出版信息

Fungal Genet Biol. 2011 Jun;48(6):641-9. doi: 10.1016/j.fgb.2010.11.013. Epub 2010 Dec 10.

Abstract

Heterotrimeric Gα protein Pga1 of Penicillium chrysogenum controls vegetative growth, conidiation and secondary metabolite production. In this work we studied the role of Pga1 in spore germination and resistance to different stress conditions. Strains G203R-T (expressing the dominant inactivating pga1(G203R) allele) and Δpga1 (deleted pga1) showed a delayed and asynchronic germination pattern, and a decrease in the percentage of germination, which occurred in only 70-80% of the total conidia. In contrast, in strains expressing the dominant activating pga1(G42R) allele, germination occurred at earlier times and in 100% of conidia. In addition, strains with the pga1(G42R) allele were able to bypass the carbon source (glucose or sucrose) requirement for germination in about 64% of conidia. Thus Pga1 plays an important, but not essential, role in germination, mediating carbon source sensing. Regulation of germination by Pga1 is probably mediated by cAMP, as intracellular levels of this secondary messenger undergo a peak before the onset of germination only in strains with an active Pga1. Pga1 activity is also a determinant factor in the resistance to different stress conditions. Absence or inactivation of Pga1 allow growth on SDS-containing minimal medium, increase resistance of conidia to thermal and oxidative stress, and increase resistance of vegetative mycelium to thermal and osmotic stress. In contrast, constitutive activation of Pga1 causes a decrease in the resistance of conidia to thermal stress and of vegetative mycelium to thermal and osmotic stress. Together with our previously reported results, we show in this work that Pga1 plays a central role in the regulation of the whole growth-developmental program of this biotechnologically important fungus.

摘要

青霉异三聚体 Gα 蛋白 Pga1 控制着营养生长、分生孢子形成和次生代谢产物的产生。在这项工作中,我们研究了 Pga1 在孢子萌发和抵抗不同胁迫条件中的作用。表达显性失活 pga1(G203R)等位基因的 G203R-T 菌株和 Δpga1(缺失 pga1)菌株表现出延迟和不同步的萌发模式,并且萌发的孢子比例下降,只有总孢子的 70-80%发生萌发。相比之下,表达显性激活 pga1(G42R)等位基因的菌株在更早的时间和 100%的孢子中发生萌发。此外,具有 pga1(G42R)等位基因的菌株能够绕过孢子萌发对碳源(葡萄糖或蔗糖)的需求,大约有 64%的孢子能够绕过碳源需求。因此,Pga1 在萌发过程中发挥着重要但非必需的作用,介导对碳源的感知。Pga1 对萌发的调控可能是通过 cAMP 介导的,因为只有在具有活性 Pga1 的菌株中,这种二级信使的细胞内水平才会在萌发开始前出现峰值。Pga1 的活性也是抵抗不同胁迫条件的决定因素。Pga1 的缺失或失活允许在含有 SDS 的最小培养基中生长,增加分生孢子对热和氧化应激的抗性,并增加营养菌丝对热和渗透胁迫的抗性。相比之下,Pga1 的组成性激活导致分生孢子对热应激和营养菌丝对热和渗透胁迫的抗性降低。结合我们之前的研究结果,我们在这项工作中表明,Pga1 在调节这种具有生物技术重要性的真菌的整个生长-发育程序中发挥着核心作用。

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