Deletion and functional studies of SER5 gene in Cryptococcus neoformans

Abstract

Abstract: Objective To construct SER5 gene deletion and reconstruction mutants for Cryptococcus neoformans and study its functions. Methods Knocking-out and reconstruction the SER5 gene were performed in strain H99, and the function of Ser5 were analyzed by in vitro stress response experiment, melanin induction experiment, capsule induction experiment, urease determination, growth curve determination, and yeast two-hybrid experiment. Results Gene deletion and reconstruction mutants of SER5 were successfully constructed. ser5Δ had no significant effect on the in vitro stress, capsule, urea decomposition and growth of C.neoformans, and it had a significant reduction in the melanin synthesis of C. neoformans.19 genes were screened by yeast two-hybrid experiment that the coding proteins may interact with Ser5. Conclusion Ser5 affected the melanin production of C. neoformans, and it might interact with endoplasmic reticulum protein, protein YOP1, vesicle-associated membrane protein 7, inorganic phosphate transporter pho88, GDP-mannose transporter 1, vesicle transport through interaction with t-SNAREs 1, suggesting that Ser5 might be involved in the synthesis or transport of melanin in C. neoformans.

Key words: Cryptococcus neoformans, serine protease, melanin, function

CLC Number:

R379.5

Tong Shuangli, WU Jie, LI Shasha, SANG Junjun. Deletion and functional studies of SER5 gene in Cryptococcus neoformans[J]. Chinese Journal of Mycology, 2021, 16(5): 335-340.

References

[1] Maziarz EK, Perfect JR. Cryptococcosis[J]. Infect Dis Clin North Am, 2016, 30(1):179-206.
[2] Rajasingham R, Smith RM, Park BJ, et al. Global burden of disease of HIV-associated cryptococcal meningitis:an updated analysis[J]. Lancet Infect Dis, 2017, 17(8):873-881.
[3] Kronstad JW, Attarian R, Cadieux B, et al. Expanding fungal pathogenesis:Cryptococcus breaks out of the opportunistic box[J]. Nat Rev Microbiol, 2011, 9(3):193-203.
[4] Yike I. Fungal proteases and their pathophysiological effects[J]. Mycopathologia, 2011, 171(5):299-323.
[5] Campbell LT, Simonin AR, Chen C, et al. Cryptococcus strains with different pathogenic potentials have diverse protein secretomes[J]. Eukaryot Cell, 2015, 14(6):554-563.
[6] Monod M, Capoccia S, Lechenne B, et al. Secreted proteases from pathogenic fungi[J]. Int J Med Microbiol, 2002, 292(5-6):405-419.
[7] Vu K, Tham R, Uhrig JP, et al. Cleavage of human fibronectin and other basement membrane-associated proteins by a Cryptococcus neoformans serine proteinase[J]. Microb Pathog, 2003, 34(2):65-71.
[8] Xu CY, Zhu HM, Wu JH, et al. Invasion of the central nervous system by Cryptococcus neoformans requires a secreted fungal metalloprotease[J]. MBio, 2014, 5(3):e01101-01114.
[9] Xu CY, Zhu HM, Wu JH, et al. Increased permeability of blood-brain barrier is mediated by serine protease during Cryptococcus meningitis[J]. J Int Med Res, 2014, 42(1):85-92.
[10] Chen Y, Toffaletti DL, Tenor JL, et al. The Cryptococcus neoformans transcriptome at the site of human meningitis[J]. MBio, 2014, 5(1):e01087-01013.
[11] Nosanchuk JD, Stark RE, Casadevall A. Fungal melanin:What do we know about structure[J]? Front Microbiol, 2015, 6(1):1463.
[12] Cordero R, Camacho E, Casadevall A. Melanization in Cryptococcus neoformans requires complex regulation[J]. mBio, 2020, 11(1):e03313-19
[13] Lee D, Jang EH, Lee M, et al. Unraveling melanin biosynthesis and signaling networks in Cryptococcus neoformans[J]. mBio, 2019, 10(5):e02267-19
[14] Kristan K, Rižner TL. Steroid-transforming enzymes in fungi[J]. J Steroid Biochem Mol Biol, 2012, 129(1-2):79-91.
[15] James AW, Nachiappan V. Phosphate transporter mediated lipid accumulation in Saccharomyces cerevisiae under phosphate starvation conditions[J]. Bioresour Technol, 2014, 151(1):100-105.
[16] Wang ZA, Griffith CL, Skowyra ML, et al. Cryptococcus neoformans dual GDP-mannose transporters and their role in biology and virulence[J]. Eukaryot Cell, 2014, 13(6):832-842.
[17] Ngamskulrungroj P, Chang Y, Hansen B, et al. Cryptococcus neoformans Yop1, an endoplasmic reticulum curvature-stabilizing protein, participates with Sey1 in influencing fluconazole-induced disomy formation[J]. FEMS Yeast Res, 2012, 12(7):748-754.