CaIPF14030 negatively modulates intracellular ATP levels during the development of azole resistance in Candida albicans
CaIPF14030 negatively modulates intracellular ATP levels during the development of azole resistance in Candida albicans
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CaIPF14030 在白色念珠菌耐药性发展过程中负向调节细胞内 ATP 水平
DOI：
10.1038/aps.2010.232
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发表时间：
2011-04-01
影响因子：
8.2
通讯作者：
Gu, Jun
中科院分区：
文献类型：
--
作者：
Jia, Xin-ming;Wang, Ying;Gu, Jun
目的：唑类药物尤其是氟康唑的广泛和重复使用导致白色念珠菌对唑类药物的耐药性迅速发展。我们研究了CaIPF14030在白色念珠菌唑类抗性发展过程中的作用。方法：通过定量RT-PCR测量CaIPF14030的表达，并通过hisG-URA3-hisG（URA-blaster）方法破坏CaIPF14030。采用点滴法检测白色念珠菌对唑类药物的敏感性，并通过光度计测定细胞内ATP浓度。结果：白色念珠菌中CaIPF14030的表达以钙调神经磷酸酶依赖性方式被Ca 2+ 上调，并且在逐步获得唑类抗性的过程中该蛋白过度表达。然而，CaIPF14030 的破坏或异位过度表达并不影响白色念珠菌对唑类的敏感性。最后，我们证明CaIPF14030的破坏显着增加了白色念珠菌的细胞内ATP水平，而过表达则显着降低了白色念珠菌的细胞内ATP水平。结论：CaIPF14030可能在白色念珠菌的唑类抗性发展过程中负向调节细胞内ATP水平。
Aim:Widespread and repeated use of azoles, particularly fluconazole, has led to the rapid development of azole resistance in Candida albicans. We investigated the role of CaIPF14030 during the development of azole resistance in C albicans.Methods:The expression of CaIPF14030 was measured by quantitative RT-PCR, and CaIPF14030 was disrupted by the hisG-URA3-hisG (URA-blaster) method. The sensitivity of C albicans to azoles was examined using a spot assay, and the intracellular ATP concentrations were measured by a luminometer.Results:CaIPF14030 expression in C albicans was up-regulated by Ca 2+ in a calcineurin-dependent manner, and the protein was overexpressed during the stepwise acquisition of azole resistance. However, disruption or ectopic overexpression of CaIPF14030 did not affect the sensitivity of C albicans to azoles. Finally, we demonstrated that disruption of CaIPF14030 significantly increased intracellular ATP levels, and overexpression significantly decreased intracellular ATP levels in C albicans.Conclusion:CaIPF14030 may negatively modulate intracellular ATP levels during the development of azole resistance in C albicans.