Using Galleria mellonella model to investigate the role of SMT3 gene in regulating pathogenicity of Candida albicans

Abstract

Abstract: Objective To investigate the role of SMT3 gene in regulating pathogenicity of Candida albicans. Methods To delete the SMT3 gene, a transient CRISPR/Cas9 system was used. The genotype of the SMT3 deletion strain was checked by colony PCR. Invasive growth and biofilm formation of the SMT3 deletion strain were studied. To check the pathogenicity of C. albicans cells, wild-type and the SMT3 deletion strain were injected into the larvae of Galleria mellonella. The survival rate of infected larvae was checked daily and analyzed. As well, the morphology of C. albicans in the larvae was checked using a PAS staining. Moreover, the damaging effect of the SMT3 deletion strain on the hemolymph of G. mellonella was investigated. Results The SMT3 homozygous deletion strain was successfully constructed. Deletion of SMT3 gene significantly increased the invasive growth on solid media, and also elevated the ability of biofilm formation. Deleting SMT3 decreased the survival of infected G. mellonella, An average survival of 3.5 days in this group is significantly lower than the wild type group (8.5 days). In addition, deleting SMT3 caused a significant decrease in total hemolymph of infected larvae, (1.4×10⁷ cells per mL). Conclusion SMT3 gene plays important roles in regulating pathogenicity of C. albicans in G. mellonella model.

Key words: Candida albicans, SMT3 gene, Galleria mellonella, pathogenicity

CLC Number:

R379.4

HU Jing, LI Aihong, FENG Jinrong. Using Galleria mellonella model to investigate the role of SMT3 gene in regulating pathogenicity of Candida albicans[J]. Chinese Journal of Mycology, 2022, 17(2): 103-108.

References

[1] FIDEL P L Jr. Candida-host interactions in HIV disease:Implications for oropharyngeal candidiasis[J]. Adv Dent Res, 2011, 23(1):45-49.
[2] AZIE N, NEOFYTOS D, PFALLER M, et al. The PATH (Prospective Antifungal Therapy) Alliance^® registry and invasive fungal infections:update 2012[J]. Diagn Microbiol Infect Dis, 2012, 73(4):293-300.
[3] MAYER F L, WILSON D, HUBE B. Candida albicans pathogenicity mechanisms[J]. Virulence, 2013, 4(2):119-128.
[4] TAKAHASHI Y, KIKUCHI Y. Cytoplasmic sumoylation by PIAS-type Siz1-SUMO ligase[J]. Cell Cycle, 2008, 7(12), 1738-1744.
[5] CHENG C, LO Y, LIANG S, et al. SUMO modifications control assembly of synaptonemal complex and polycomplex in meiosis of Saccharomyces cerevisiae[J].Genes Dev, 2006, 20(15):2067-2081.
[6] YAN M, NIE X, WANG H, et al. SUMOylation of Wor1 by a novel SUMO E3 ligase controls cell fate in Candida albicans[J]. Mol Microbiol, 2015, 98(1):69-89.
[7] LEACH M D, STEAD D A, ARGO E, et al. Identification of sumoylation targets, combined with inactivation of SMT3, reveals the impact of sumoylation upon growth, morphology, and stress resistance in the pathogen Candida albicans[J]. Mol Biol Cell, 2011, 22(5):525-714.
[8] HARDING C R, SCHROEDER G N, COLLINS J W, et al. Use of Galleria mellonella as a model organism to study Legionella pneumophila infection[J]. J Vis Exp, 2013, (81):50964. DOI:10.3791/50964.
[9] SCORZONI L, DE LUCAS M P, MESA-ARANGO A C, et al. Antifungal efficacy during Candida krusei infection in non-conventional models correlates with the yeast in vitro susceptibility profile[J]. PLoS ONE, 2013, 8(3):e60047.
[10] KIM C H, SHIN Y P, NOH M Y, et al. An insect multiligand recognition protein functions as an opsonin for the phagocytosis of microorganisms[J]. J Biol Chem 2010, 285(33), 25243-25250.
[11] 赵望, 孙毅, 曾同祥. 大蜡螟作为病原真菌动物实验模型的相关研究[J]. 中国真菌学杂志, 2020, 15(4):248-252.
[12] 冯佳, 胡婧, 单爱迪, 等. 利用CRISPR/Cas9系统快速构建白念珠菌RAD23高表达菌株的研究[J]. 中国病原生物学杂志, 2019, 14(9):1019-1023.
[13] 单爱狄, 冯金荣. 白念珠菌SUMO结合酶UBC9基因缺失株的构建及其在形态调控中的作用研究[J]. 中国病原生物学杂志, 2020, 15(7):800-804.
[14] ISLAM A, TEBBJI F, MALLICK J, et al. Mms21:A putative SUMO E3 ligase in Candida albicans that negatively regulates invasiveness and filamentation, and is required for the genotoxic and cellular stress response[J]. Genetics, 2019, 211(2):579-595.
[15] HUI C, XUE D, BIAO R, et al. The regulation of hyphae growth in Candida albicans[J]. Virulence, 2020, 11(1):337-348.
[16] FALLON A M, SUN D X. Exploration of mosquito immunity using cells in culture[J]. Insect Biochem Mol Biol, 2001, 31(3):263-278.
[17] SHEEHAN G, KAVANAGH K.Proteomic analysis of the responses of Candida albicans during infection of Galleria mellonella larvae[J]. J FUNGI, 2019, 5(1):7.