Alkaline stress triggers an immediate calcium fluctuation in Candida albicans mediated by Rim101p and Crz1p transcription factors

In the human fungal pathogen Candida albicans, environmental pH has profound effects on morphogenesis and response to extracellular pH is clearly relevant to the pathogenicity of this fungus. Yeast cells have evolved a complex network of mechanisms in response to the environmental pH and they often require the integration of the Rim101 and calcineurin/Crz1 signaling pathways. Ca(2+) burst is a common cellular response when cells are exposed to environmental stresses; therefore, in this study, we asked whether it follows the same case under alkaline stress and whether this calcium change is regulated by Rim101p and Crz1p. We confirmed the calcium influx was activated by KOH stimuli using a flow cytometry-based method, but it was obviously abolished in cells lacking MID1 or CCH1. We also found that alkaline pH-induced activation of the PHO89 promoter was blocked without the same gene; moreover, the response was Crz1p- and Rim101p-dependent. Finally, we investigated the regulation role of Rim101p and Crz1p in calcium influx through MID1, CCH1 and YVC1 genes, which were all downregulated in rim101Δ/Δ and crz1Δ/Δ mutants. The important role of calcium influx in the alkaline stress response and its regulation suggested a potential integration effect of Rim101 and Crz1 signaling pathways in C. albicans.     

几种白念珠菌高铁还原酶基因的功能研究

【目的】高铁还原酶在白念珠菌铁离子获得过程中发挥着重要作用。通过研究高铁还原酶基因的功能和表达调控,揭示其不同的环境应答策略。【方法】通过Northern杂交的方法分析不同低铁环境对高铁还原酶基因表达水平的影响。构建高铁还原酶基因缺失菌株,探究基因缺失后对细胞表面高铁还原酶活力和生长状况的影响。利用激光共聚焦显微镜观察Frp1蛋白的细胞学定位。【结果】酸性条件显著上调FRE10基因的表达水平,而碱性条件能显著提高FRE2基因的表达量。在酸性条件下,FRE10基因的缺失会显著地下调细胞表面高铁还原酶活力。在碱性条件下,fre2Δ/Δ缺失菌株表现出严重缺陷的生长能力和显著降低的表面高铁还原酶活力。细胞学定位实验发现Frp1蛋白位于液泡中。【结论】FRE2和FRE10基因的表达模式主要是酸碱依赖性的。Fre2是碱性条件下高铁还原酶活力的主要贡献者。Frp1蛋白位于液泡中,在液泡内储存铁的活化和转运过程中可能发挥重要作用。     

白念珠菌唑类药物耐药相关转录因子研究进展

近年来白念珠菌的感染率呈逐年上升趋势,随着唑类药物的广泛应用,耐药菌株不断增多,已成为临床治疗的一大难题.白念珠菌的耐药机制主要与ERG 11基因的突变和过表达、药物外排泵相关基因表达增多及生物膜的形成等有关,由于转录因子是耐药基因表达的关键调节因子,关于锌簇转录因子与耐药关系的研究越来越多,如TAC 1、MRR 1、MRR 2、UPC 2、NDT 80等,其点突变可引起某些耐药基因的过表达而介导耐药,该领域研究已成为热点,该文就此研究进展做一概述.     

Thermotolerance and the heat-shock response in Candida albicans

At elevated temperatures, yeast cells of Candida albicans synthesized nine heat-shock proteins (HSPs) with apparent molecular masses of 98, 85, 81, 76, 72, 54, 34, 26 and 18 kDa. The optimum temperature for the heat-shock response was 45 degrees C although HSPs were detected throughout the range 41-46 degrees C. Protein synthesis was not observed in cells kept at 48 degrees C. Yeast cells survived exposure to an otherwise lethal temperature of 55 degrees C when they had previously been exposed to 45 degrees C. The thermotolerance induced during incubation at 45 degrees C required protein synthesis, since protection was markedly reduced by trichodermin. Mercury ions induced a set of three stress proteins, one of which corresponded in size to an HSP, and cadmium ions evoked one stress protein seemingly unrelated to the HSPs observed after temperature shift.     

Deletion of the copper transporter CaCCC2 reveals two distinct pathways for iron acquisition in Candida albicans

Efficient iron acquisition is an essential requirement for growth of pathogenic organisms in the iron-poor host environment. In Saccharomyces cerevisiae, high-affinity iron import depends on the multicopper ferroxidase ScFet3. ScFet3 biogenesis in the trans-Golgi compartment requires a copper-transporting P-type ATPase, ScCcc2. Here, we describe the isolation by functional complementation of a Ccc2 homologue from the pathogenic yeast Candida albicans. CaCcc2 is functionally distinct from a previously described C. albicans copper-transporting P-type ATPase, CaCrp1, which appears to be specifically involved in copper detoxification. Regulation of CaCCC2 and the phenotype of the homozygous CaCCC2 deletion indicate that it is required for high-affinity iron import, making it the bona fide CCC2 homologue of C. albicans. Remarkably, in a mouse model of systemic infection, the Caccc2Delta strain displayed robust proliferation and no significant reduction in pathogenicity, suggesting the existence of alternative mechanisms of iron uptake from host tissues. We identify haemin and haemoglobin as potential iron sources that can be used by C. albicans in a CaCcc2-independent manner.     

植物缺铁应答bHLH转录因子研究进展

铁是植物生命活动必需的微量元素之一,土壤中有效铁含量较低,易导致植物缺铁。bHLH转录因子家族多个成员参与植物缺铁响应,发挥重要的调控作用。为深入了解植物对缺铁的反应机制,文中对植物缺铁胁迫应答的bHLH转录因子的结构、分类和功能及其调控机制、介导的缺铁胁迫信号通路进行综述,为应用bHLH转录因子培育缺铁耐受作物或富铁作物提供理论依据和设计策略。