Proteomic and Transcriptomic Analysis of Aspergillus fumigatus on Exposure to Amphotericin B
Proteomic and Transcriptomic Analysis of Aspergillus fumigatus on Exposure to Amphotericin B
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DOI：
10.1128/aac.01431-07
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发表时间：
2008-12-01
影响因子：
4.9
通讯作者：
Sarma, Puranam Usha
中科院分区：
文献类型：
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作者：
Gautam, Poonam;Shankar, Jata;Sarma, Puranam Usha
两性霉素B（AMB）是目前应用最广泛的多烯类抗真菌药物，用于治疗包括侵袭性曲霉病在内的全身真菌感染。通过基因组学和蛋白质组学方法研究AMB在烟曲霉中的作用靶点是我们的目标。在转录组学分析中，共发现295个基因差异表达（165个上调和130个下调），包括许多涉及麦角固醇途径，细胞应激蛋白，细胞壁蛋白，转运蛋白和假设蛋白。A. fumigatus单独或A.用AMB处理的烟曲霉显示85种蛋白质的差异表达水平（76种上调，9种下调）。其中48个被确定为高度可信，属于上述类别。本文首次报道了Rho-GDP解离抑制因子（Rho-GDI）、分泌途径GDI、网格蛋白、Sec 31（胞外复合物的一个亚基）和RAB GTptYpt 51在抗真菌药物作用下的差异表达水平，并可能代表了A.烟曲霉菌AMB。其中一些基因的表达通过实时逆转录-PCR验证。AMB反应基因/蛋白在A.烟曲霉可进一步用于新药开发。
Amphotericin B (AMB) is the most widely used polyene antifungal drug for the treatment of systemic fungal infections, including invasive aspergillosis. It has been our aim to understand the molecular targets of AMB in Aspergillus fumigatus by genomic and proteomic approaches. In transcriptomic analysis, a total of 295 genes were found to be differentially expressed (165 upregulated and 130 downregulated), including many involving the ergosterol pathway, cell stress proteins, cell wall proteins, transport proteins, and hypothetical proteins. Proteomic profiles of A. fumigatus alone or A. fumigatus treated with AMB showed differential expression levels for 85 proteins (76 upregulated and 9 downregulated). Forty-eight of them were identified with high confidence and belonged to the above-mentioned categories. Differential expression levels for Rho-GDP dissociation inhibitor (Rho-GDI), secretory-pathway GDI, clathrin, Sec 31 (a subunit of the exocyst complex), and RAB GTPase Ypt51 in response to an antifungal drug are reported here for the first time and may represent a specific response of A. fumigatus to AMB. The expression of some of these genes was validated by real-time reverse transcription-PCR. The AMB responsive genes/proteins observed to be differentially expressed in A. fumigatus may be further explored for novel drug development.