烟曲霉Cyp51A和Cyp51B生物学功能差异性分析

中国真菌学杂志　 2021, Vol. 16 　Issue (6): 373-378.

论著上一篇

烟曲霉Cyp51A和Cyp51B生物学功能差异性分析

房凌旭¹, 梁海¹, 王鹏远¹, 卢中一², 陈芳艳³, 韩黎³

1. 深圳市前海蛇口自贸区医院口腔科, 深圳 518067;
2. 深圳大学高等研究院深圳市海洋微生物组工程重点实验室, 深圳 518060;
3. 中国人民解放军疾病预防控制中心消毒与感染控制科, 北京 100071

收稿日期:2020-11-24 发布日期:2021-12-29
通讯作者: 韩黎,E-mail:hanlicdc@163.com;陈芳艳,E-mail:chenfangyan1986@163.com E-mail:hanlicdc@163.com;chenfangyan1986@163.com
作者简介:房凌旭,女(汉族),硕士,主治医师.E-mail:zyl531@yeah.net
基金资助:
国家自然科学基金（81971914）；广东省基础与应用基础研究基金（2019A1515110089）；深圳市南山区卫生科技项目（2020057）

Analysis of functional differentiation between Cyp51A and Cyp51B of Aspergillus fumigatus

FANG Lingxu¹, LIANG Hai¹, WANG Pengyuan¹, LU Zhongyi², CHEN Fangyan³, HAN Li³

1. Department of stomatology, Shenzhen Qianhai Shekou Free Trade Zone Hospital, Shenzhen 518067, China;
2. Shenzhen Key Laboratory of Marine Microbiome Engineering, Institute for Advanced Study, Shenzhen University, Shenzhen 518060, China;
3. Department for Disinfection & Infection Control, Chinese PLA Center for Disease Control and Prevention, Beijing 10071, China

Received:2020-11-24 Published:2021-12-29

摘要/Abstract

摘要： 目的研究烟曲霉Cyp51A和Cyp51B的功能差异。方法利用系统发生分析研究烟曲霉Cyp51A和Cyp51B的进化关系,分别构建烟曲霉Cyp51A和Cyp51B编码基因的敲除菌株Δcyp51A和Δcyp51B,研究Δcyp51A和Δcyp51B在高温耐受性、伏立康唑敏感性和对大蜡螟幼虫致病力方面的差异性。结果系统发生分析显示,烟曲霉Cyp51A和Cyp51B进化距离较远。Δcyp51B对50℃的耐受性低于Δcyp51A和野生株。Δcyp51A对伏立康唑耐受水平下降程度明显高于Δcyp51B。Δcyp51B和Δcyp51A对大蜡螟幼虫致病力与野生株相比无显著性差异。结论 Cyp51A和Cyp51B在烟曲霉高温耐受性和唑类药物耐受性中有功能差异。

关键词: 烟曲霉, 羊毛甾醇14-α-脱甲基酶, 功能分化

Abstract: Objective The functional differentiation of Aspergillus fumigatus Cyp51A and Cyp51B was characterized in this study. Method The phylogenetic tree of Cyp51A and Cyp51B was constructed by IQ-tree. The Aspergillus fumigatus Δcyp51A and Δcyp51B mutants were constructed by homologous recombination technology, respectively. The high temperature tolerance and voriconazole susceptibility of both Δcyp51A and Δcyp51B mutants were determined. Moreover, the virulence of Δcyp51A and Δcyp51B mutants was investigated in the Galleria mellonella infection model. Result Phylogenetic analysis revealed that the Cyp51Bs formed a cluster that was separated from Cyp51As. The Δcyp51B mutant showed reduced tolerance to 50 °C when compared to the Δcyp51A and wild-type strain. The Δcyp51A mutant manifested significantly reduced tolerance to voriconazole when compared to that in Δcyp51B mutant. Additionally, there was no statistically significant difference among the Δcyp51A mutant, Δcyp51B mutant, and the wild-type strain. Conclusion The Cyp51A held different functional roles in high temperature tolerance and azole susceptibility of A. fumigatus when compared with Cyp51B.

Key words: Aspergillus fumigatus, Cyp51, functional differentiation

中图分类号:

R379.6

房凌旭, 梁海, 王鹏远, 卢中一, 陈芳艳, 韩黎. 烟曲霉Cyp51A和Cyp51B生物学功能差异性分析[J]. 中国真菌学杂志, 2021, 16(6): 373-378.

FANG Lingxu, LIANG Hai, WANG Pengyuan, LU Zhongyi, CHEN Fangyan, HAN Li. Analysis of functional differentiation between Cyp51A and Cyp51B of Aspergillus fumigatus[J]. Chinese Journal of Mycology, 2021, 16(6): 373-378.

参考文献

[1] KKOUSHA M, TADI R, SOUBANI A O. Pulmonary aspergillosis: a clinical review[J]. Eur Respir Rev, 2011, 20(121): 156-174.
[2] DENNING D W, PLEUVRY A, COLE D C. Global burden of allergic bronchopulmonary aspergillosis with asthma and its complication chronic pulmonary aspergillosis in adults[J]. Med Mycol, 2013, 51(4): 361-370.
[3] BERGER S, El CHAZLI Y, BABU A F, et al. Azole resistance in Aspergillus fumigatus: a consequence of antifungal use in agriculture[J]? Front Microbiol, 2017, 8: 1024. DOI: 10.3389/fmicb.2017.01024.
[4] KRISHNAN-NATESAN S, CHANDRASEKAR P H, ALANGADEN G J, et al. Molecular characterisation of cyp51A and cyp51B genes coding for P45014alpha-lanosterol demethylases A (CYP51Ap) and B (CYP51Bp) from voriconazole-resistant laboratory isolates of Aspergillus flavus[J]. Int J Antimicrob Agents, 2008, 32(6): 519-524.
[5] CHOWDHARY A, SHARMA C, HAGEN F, et al. Exploring azole antifungal drug resistance in Aspergillus fumigatus with special reference to resistance mechanisms[J]. Future Microbiol, 2014, 9(5): 697-711.
[6] CAMPS S M, RIJS A J, KLAASSEN C H, et al. Molecular epidemiology of Aspergillus fumigatus isolates harboring the TR34/L98H azole resistance mechanism[J]. J Clin Microbiol, 2012, 50(8): 2674-2680.
[7] ALVAREZ-MORENO C, LAVERGNE R A, HAGEN F, et al. Azole-resistant Aspergillus fumigatus harboring TR34/L98H, TR46/Y121F/T289A and TR53 mutations related to flower fields in Colombia[J]. Sci Rep, 2017, 7: 45631. DOI: 10.1038/srep45631.
[8] MAGUIRE S L, WANG C, HOLLAND L M, et al. Zinc finger transcription factors displaced SREBP proteins as the major sterol regulators during Saccharomycotina evolution[J]. PLoS Genet, 2014, 10(1): e1004076.
[9] DHINGRA S, KOWALSKI C H, THAMMAHONG A, et al. RbdB, a rhomboid protease critical for SREBP activation and virulence in Aspergillus fumigatus[J]. mSphere, 2016, 1(2): e00035-16.
[10] BRILLOWSKA-DABROWSKA A, MROCZYNSKA M, NAWROT U, et al. Examination of cyp51A and cyp51B expression level of the first polish azole resistant clinical Aspergillus fumigatus isolate[J]. Acta Biochim Pol, 2015, 62(4): 837-839.
[11] CHEN S, WANG Y, SCHNABEL G, et al. Inherent resistance to 14alpha-demethylation inhibitor fungicides in Colletotrichum truncatum is likely linked to CYP51A and/or CYP51B gene variants[J]. Phytopathology, 2018, 108(11): 1263-1275.
[12] LIU X, YU F, SCHNABEL G, et al. Paralogous cyp51 genes in Fusarium graminearum mediate differential sensitivity to sterol demethylation inhibitors[J]. Fungal Genet Biol, 2011, 48(2): 113-123.
[13] LU Z, JIA X, CHEN Y, et al. Identification and characterization of key charged residues in the cofilin protein involved in azole susceptibility, apoptosis, and virulence of Aspergillus fumigatus[J]. Antimicrobial agents and chemotherapy, 2018, 62(5): e01659-01617.
[14] EDGAR R C. MUSCLE: a multiple sequence alignment method with reduced time and space complexity[J]. BMC Bioinformatics, 2004, 5(1): 1-19.
[15] CAPELLA-GUTIERREZ S, SILLA-MARTINEZ J M, GABALDON T. TrimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses[J]. Bioinformatics, 2009, 25(15): 1972-1973.
[16] NGUYEN L T, SCHMIDT H A, VON HAESELER A, et al. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies[J]. Mol Biol Evol, 2015, 32(1): 268-274.
[17] BECHER R, WIRSEL S G. Fungal cytochrome P450 sterol 14alpha-demethylase (CYP51) and azole resistance in plant and human pathogens[J]. Appl Microbiol Biotechnol, 2012, 95(4): 825-840.
[18] GARCIA-RUBIO R, ALCAZAR-FUOLI L, MONTEIRO M C, et al. Insight into the significance of Aspergillus fumigatus cyp51A polymorphisms[J]. Antimicrob Agents Chemother, 2018, 62(6): e00241-18.
[19] WARRILOW A G, MELO N, MARTEL C M, et al. Expression, purification, and characterization of Aspergillus fumigatus sterol 14-alpha demethylase (CYP51) isoenzymes A and B[J]. Antimicrob Agents Chemother, 2010, 54(10): 4225-4234.
[20] CHEN Y, LU Z, ZHAO J, et al. Epidemiology and molecular characterizations of azole resistance in clinical and environmental Aspergillus fumigatus isolates from China[J]. Antimicrob Agents Chemother, 2016, 60(10): 5878-5884.
[21] ZHANG J, LI L, LV Q, et al. The fungal CYP51s: Their functions, structures, related drug resistance, and inhibitors[J]. Front Microbiol, 2019, 10: 691. DOI: 10.3389/fmicb.2019,00691.
[22] SORIANI F M, MALAVAZI I, DA SILVA FERREIRA M E, et al. Functional characterization of the Aspergillus fumigatus CRZ1 homologue, CrzA[J]. Mol Microbiol, 2008, 67(6): 1274-1291.
[23] VERWEIJ P E, ZHANG J, DEBETS A J M, et al. In-host adaptation and acquired triazole resistance in Aspergillus fumigatus: a dilemma for clinical management[J]. Lancet Infect Dis, 2016, 16(11): e251-e260.
[24] LIU G, CHEN Y, FAERGEMAN N J, et al. Elimination of the last reactions in ergosterol biosynthesis alters the resistance of Saccharomyces cerevisiae to multiple stresses[J]. FEMS Yeast Res, 2017, 17(6). DOI: 10.1093/femsyr/fox063.
[25] BHABHRA R, ASKEW D S. Thermotolerance and virulence of Aspergillus fumigatus: role of the fungal nucleolus[J]. Med Mycol, 2005, 43(Suppl 1): S87-93.

烟曲霉Cyp51A和Cyp51B生物学功能差异性分析

Analysis of functional differentiation between Cyp51A and Cyp51B of Aspergillus fumigatus

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