Abstract
Background
The Candida glabrata does not develop into a pathogenic hiphal form; however, it has become the second most common pathogen of fungal infections in humans, partly because of its adhesion ability and virulence.
Objectives
The present study aimed to determine whether Flo8, a transcription factor that plays an important role in the virulence and drug resistance in Candida albicans, has a similar role in C. glabrata.
Methods
We constructed FLO8 null strains of a C. glabrata standard strain and eight clinical strains from different sources, and a FLO8 complemented strain. Real-time quantitative PCR, biofilm formation assays, hydrophobicity tests, adhesion tests, Caenorhabditis elegans survival assay, and drug-susceptibility were then performed.
Results
Compared with the wild-type strains, the biofilm formation, hydrophobicity, adhesion, and virulence of the FLO8-deficient strains decreased, accompanied by decreased expression of EPA1, EPA6, and EPA7. On the other hand, it showed no changes in antifungal drug resistance, although the expression levels of CDR1, CDR2, and SNQ2 increased after FLO8 deletion.
Conclusions
These results indicated that Flo8 is involved in the adhesion and virulence of C. glabrata, with FLO8 deletion leading to decreased expression of EPA1, EPA6, and EPA7 and decreased biofilm formation, hydrophobicity, adhesion, and virulence.
Similar content being viewed by others
References
Valotteau C, Prystopiuk V, Cormack BP, Dufrene YF (2019) Atomic force microscopy demonstrates that candida glabrata uses three Epa proteins to mediate adhesion to abiotic surfaces. Msphere 4(3):e00277-e319. https://doi.org/10.1128/mSphere.00277-19
Goncalves B, Ferreira C, Alves CT, Henriques M, Azeredo J, Silva S (2016) Vulvovaginal candidiasis: epidemiology, microbiology and risk factors. Crit Rev Microbiol 42(6):905–927. https://doi.org/10.3109/1040841x.2015.1091805
Guinea J (2014) Global trends in the distribution of Candida species causing candidemia. Clin Microbiol Infect 20(Suppl 6):5–10. https://doi.org/10.1111/1469-0691.12539
Andes DR, Safdar N, Baddley JW, Alexander B, Brumble L, Freifeld A, Hadley S, Herwaldt L, Kauffman C, Lyon GM, Morrison V, Patterson T, Perl T, Walker R, Hess T, Chiller T, Pappas PG, Investigators T (2016) The epidemiology and outcomes of invasive Candida infections among organ transplant recipients in the United States: results of the Transplant-Associated Infection Surveillance Network (TRANSNET). Transpl Infect Dis 18(6):921–931. https://doi.org/10.1111/tid.12613
Cleveland AA, Harrison LH, Farley MM, Hollick R, Stein B, Chiller TM, Lockhart SR, Park BJ (2015) Declining incidence of candidemia and the shifting epidemiology of candida resistance in two US metropolitan areas, 2008–2013: results from population-based surveillance. PLoS ONE.10(3). https://doi.org/10.1371/journal.pone.0120452
Krcmery V, Barnes AJ (2002) Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance. J Hosp Infect. 50(4):243–260. https://doi.org/10.1053/jhin.2001.1151
Desai C, Mavrianos J, Chauhan N (2011) Candida glabrata Pwp7p and Aed1p are required for adherence to human endothelial cells. FEMS Yeast Res 11(7):595–601. https://doi.org/10.1111/j.1567-1364.2011.00743.x
Sinnott JTt, Cullison JP, Sweeney MP (1987) Candida (Torulopsis) glabrata. Infection control : IC. 8(8):334–336. https://doi.org/10.1017/s0195941700066443
Galocha M, Pais P, Cavalheiro M, Pereira D, Viana R, Teixeira MC (2019) Divergent approaches to virulence in C. albicans and C. glabrata: two sides of the same coin. Int J Mol Sci.20(9). https://doi.org/10.3390/ijms20092345
Rodrigues CF, Silva S, Henriques M (2014) Candida glabrata: a review of its features and resistance. Eur J Clin Microbiol Infect Dis 33(5):673–688. https://doi.org/10.1007/s10096-013-2009-3
Timmermans B, De las Penas A, Castano I, Van Dijck P (2018) Adhesins in Candida glabrata. Journal of Fungi 4(2). https://doi.org/10.3390/jof4020060
Castano I, Pan SJ, Zupancic M, Hennequin C, Dujon B, Cormack BP (2005) Telomere length control and transcriptional regulation of subtelomeric adhesins in Candida glabrata. Mol Microbiol 55(4):1246–1258. https://doi.org/10.1111/j.1365-2958.2004.04465.x
Kobayashi O, Suda H, Ohtani T, Sone H (1996) Molecular cloning and analysis of the dominant flocculation gene FLO8 from Saccharomyces cerevisiae. Mol Gen Genet 251(6):707–715. https://doi.org/10.1007/s004380050220
Cao F, Lane S, Raniga PP, Lu Y, Zhou Z, Ramon K, Chen JY, Liu HP (2006) The Flo8 transcription factor is essential for hyphal development and virulence in Candida albicans. Mol Biol Cell 17(1):295–307. https://doi.org/10.1091/mbc.E05-06-0502
Fox EP, Bui CK, Nett JE, Hartooni N, Mui MC, Andes DR, Nobile CJ, Johnson AD (2015) An expanded regulatory network temporally controls Candida albicans biofilm formation. Mol Microbiol 96(6):1226–1239. https://doi.org/10.1111/mmi.13002
Li W-J, Liu J-Y, Shi C, Zhao Y, Meng L-n, Wu F, Xiang M-J (2019) FLO8 deletion leads to azole resistance by upregulating CDR1 and CDR2 in Candida albicans. Res Microbiol 170(6–7):272–279. https://doi.org/10.1016/j.resmic.2019.08.005
Tian Y, Gao N, Ni Q, Mao Y, Dong D, Huang X, Jiang C, Li Z, Zhang L, Wang X, Peng Y, Chen C (2018) Sequence modification of the master regulator Pdr1 interferes with its transcriptional autoregulation and confers altered azole resistance in Candida glabrata. FEMS Yeast Res.18(4). https://doi.org/10.1093/femsyr/foy038
Liu J-Y, Li W-J, Shi C, Wang Y, Zhao Y, Xiang M-J (2015) Mutations in the Flo8 transcription factor contribute to virulence and phenotypic traits in Candida albicans strains. Microbiol Res 178:1–8. https://doi.org/10.1016/j.micres.2015.05.007
Castano I, Kaur R, Pan SJ, Cregg R, De Las PA, Guo NN, Biery MC, Craig NL, Cormack BP (2003) Tn7-based genome-wide random insertional mutagenesis of Candida glabrata. Genome Res 13(5):905–915. https://doi.org/10.1101/gr.848203
Wu J, Luo Q, Liu J, Chen X, Liu L (2018) Enhanced pyruvate production in Candida glabrata by overexpressing the CgAMD1 gene to improve acid tolerance. Biotechnol Lett 40(1):143–149. https://doi.org/10.1007/s10529-017-2452-9
Kottom TJ, Limper AH (2016) Evidence for a Pneumocystis carinii Flo8-like transcription factor: insights into organism adhesion. Med Microbiol Immunol 205(1):73–84. https://doi.org/10.1007/s00430-015-0428-8
Del Rio M, de la Canal L, Pinedo M, Mora-Montes HM, Regente M (2019) Effects of the binding of a Helianthus annuus lectin to Candida albicans cell wall on biofilm development and adhesion to host cells. Phytomedicine 58. https://doi.org/10.1016/j.phymed.2019.152875
Ni Q, Wang C, Tian Y, Dong D, Jiang C, Mao E, Peng Y (2018) CgPDR1 gain-of-function mutations lead to azole-resistance and increased adhesion in clinical Candida glabrata strains. Mycoses 61(7):430–440. https://doi.org/10.1111/myc.12756
Breger J, Fuchs BB, Aperis G, Moy TI, Ausubel FM, Mylonakis E (2007) Antifungal chemical compounds identified using a C-elegans pathogenicity assay. PLoS Pathog 3(2):168–178. https://doi.org/10.1371/journal.ppat.0030018
Cordeiro RdA, de Jesus Evangelista AJ, Serpa R, Colares de Andrade AR, Leite Mendes PB, Franco JdS, de Oliveira JS, de Alencar LP, Sales JA, Carneiro Camara LM, Collares Maia Castelo-Branco DdS, Nogueira Brilhante RS, Costa Sidrim JJ, Gadelha Rocha MF (2018) beta-lactam antibiotics & vancomycin increase the growth & virulence of Candida spp. Future Microbiol 13(8):869–875. https://doi.org/10.2217/fmb-2018-0019
Institute CaLS (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard. In: CLSI Document M27-A3, Clinical and Laboratory Standards Institute: Wayne.
De las Penas A, Pan SJ, Castano I, Alder J, Cregg R, Cormack BP (2003) Virulence-related surface glycoproteins in the yeast pathogen Candida glabrata are encoded in subtelomeric clusters and subject to RAP1- and SIR-dependent transcriptional silencing. Genes Dev. 17(18):2245–2258. https://doi.org/10.1101/gad.1121003
Cormack BP, Ghori N, Falkow S (1999) An adhesin of the yeast pathogen Candida glabrata mediating adherence to human epithelial cells. Science 285(5427):578–582. https://doi.org/10.1126/science.285.5427.578
Domergue R, Castano I, De las Penas A, Zupancic M, Lockatell V, Hebel JR, Johnson D, Cormack BP (2005) Nicotinic acid limitation regulates silencing of Candida adhesins during UTI. Science. 308(5723):866–870. https://doi.org/10.1126/science.1108640
Zupancic ML, Frieman M, Smith D, Alvarez RA, Cummings RD, Cormack BP (2008) Glycan microarray analysis of Candida glabrata adhesin ligand specificity. Mol Microbiol 68(3):547–559. https://doi.org/10.1111/j.1365-2958.2008.06184.x
Kraneveld EA, de Soet JJ, Deng DM, Dekker HL, de Koster CG, Klis FM, Crielaard W, de Groot PWJ (2011) Identification and differential gene expression of adhesin-like wall proteins in Candida glabrata biofilms. Mycopathologia 172(6):415–427. https://doi.org/10.1007/s11046-011-9446-2
Gomez-Molero E, de Boer AD, Dekker HL, Moreno-Martinez A, Kraneveld EA, Ichsan, Chauhan N, Weig M, de Soet JJ, de Koster CG, Bader O, de Groot PWJ (2015) Proteomic analysis of hyperadhesive Candida glabrata clinical isolates reveals a core wall proteome and differential incorporation of adhesins. FEMS Yeast Res 15(8). https://doi.org/10.1093/femsyr/fov098
Santos R, Costa C, Mil-Homens D, Romao D, de Carvalho CCCR, Pais P, Mira NP, Fialho AM, Teixeira MC (2017) The multidrug resistance transporters CgTpo1_1 and CgTpo1_2 play a role in virulence and biofilm formation in the human pathogen Candida glabrata. Cell Microbiol 19(5). https://doi.org/10.1111/cmi.12686
Kucharikova S, Tournu H, Lagrou K, Van Dijck P, Bujdakova H (2011) Detailed comparison of Candida albicans and Candida glabrata biofilms under different conditions and their susceptibility to caspofungin and anidulafungin. J Med Microbiol 60(9):1261–1269. https://doi.org/10.1099/jmm.0.032037-0
Teresa Blanco M, Sacristan B, Lucio L, Blanco J, Perez-Giraldo C, Candido Gomez-Garcia A (2010) Cell surface hydrophobicity as an indicator of other virulence factors in Candida albicans. Rev Iberoam Micol 27(4):195–199. https://doi.org/10.1016/j.riam.2010.09.001
Van Mulders SE, Christianen E, Saerens SMG, Daenen L, Verbelen PJ, Willaert R, Verstrepen KJ, Delvaux FR (2009) Phenotypic diversity of Flo protein family-mediated adhesion in Saccharomyces cerevisiae. FEMS Yeast Res 9(2):178–190. https://doi.org/10.1111/j.1567-1364.2008.00462.x
Liu H, Styles CA, Fink GR (1996) Saccharomyces cerevisiae S288C has a mutation in FLO8, a gene required for filamentous growth. Genetics 144(3):967–978
Mundy RD, Cormack B (2009) Expression of Candida glabrata adhesins after exposure to chemical preservatives. J Infect Dis 199(12):1891–1898. https://doi.org/10.1086/599120
Su C, Li Y, Lu Y, Chen J (2009) Mss11, a transcriptional activator, is required for hyphal development in Candida albicans. Eukaryot Cell 8(11):1780–1791. https://doi.org/10.1128/EC.00190-09
Kim TS, Kim HY, Yoon JH, Kang HS (2004) Recruitment of the Swi/Snf complex by Ste12-Tec1 promotes Flo8-Mss11-mediated activation of STA1 expression. Mol Cell Biol 24(21):9542–9556. https://doi.org/10.1128/MCB.24.21.9542-9556.2004
Funding
This work was supported by the National Natural Science Foundation of China [#81871706], Shanghai Municipal Health and Family Planning Commission [#201840227] and [#201740069], Natural Science Foundation of Shanghai [#15ZR1426900], the Program of Shanghai Key Specialty [#ZK2012A21], and Excellent Youth of HuangPu District of Shanghai [#RCPY1407].
Author information
Authors and Affiliations
Contributions
Conceptualization: Jun-Tao Zhao and Ming-Jie Xiang; Methodology: Jun-Tao Zhao and Jin-Yan Liu; Formal analysis and investigation: Jun-Tao Zhao, Ke-Zhi Chen, Yu-Zhu Wang, and Lu-Ling Wang; Writing—original draft preparation: Jun-Tao Zhao; Writing—review and editing: Jin-Yan Liu and Ke-Zhi Chen; Funding acquisition: Ming-Jie Xiang; Resources: Ming-Jie Xiang and Wei-Hua Li; Supervision: Ming-Jie Xiang and Jin-Yan Liu.
Corresponding author
Ethics declarations
Ethics approval
Not applicable.
Consent to participate
Not applicable.
Consent to publication
Not applicable.
Conflict of interest
The authors declare no competing interests.
Additional information
Responsible Editor: Rosana Puccia
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Authors Jun-Tao Zhao, Ke-Zhi Chen, and Jin-Yan Liu share first authorship.
Rights and permissions
About this article
Cite this article
Zhao, JT., Chen, KZ., Liu, JY. et al. FLO8 deletion leads to decreased adhesion and virulence with downregulated expression of EPA1, EPA6, and EPA7 in Candida glabrata. Braz J Microbiol 53, 727–738 (2022). https://doi.org/10.1007/s42770-022-00703-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42770-022-00703-7