Effects of polyubiquitin UBI4 downregulation on growth and pathogenesis of Cryptococcus neoformans

Abstract

Abstract: Objective To construct Cryptococcus neoformans strain with downregulated UBI4 expression and examine the effects of UBI4 downregulation on growth and pathogenesis of C.neoformans.Methods UBI4 DNA fragment was amplified and infused into linear plasmid pCH20 containing histone promoter.The fused plasmid was then transformed into C.neoformans through gene gun.The positive transformants were screened by diagnostic PCR and DNA sequencing.Various phenotype of C.neoformans reconstituted strain H99^His-UBI4 were examined.Results Cryptococcus neoformans strain with low expression of UBI4 were constructed and phenotypic assays demonstrated that down-regulated UBI4 lead to slower growth rate and enhanced susceptibility to various stresses such as H₂O₂,NO and antifungal agents in C.neoformans.Moreover,C.neoformans with down-regulated UBI4 was also attenuated dramatically in wax moth infection model.Conclusion Our study suggested that polyubiquitin UBI4 played a critical role in cell proliferation,stress response and pathogenesis in C.neoformans.Our work laid a foundation for following studies about functions and mechanisms of ubiquitin system.

Key words: Cryptococcus neoformans, polyubiquitin, pathogenesis, stress response

CLC Number:

R379.5

MENG Yun-fang, ZHOU Zhao-jing, YANG Ya-li, ZHAO Jing-yu, ZHANG Chao, FA Zhen-zong, SANG Jun-jun, FANG Wei, LIAO Wan-qing. Effects of polyubiquitin UBI4 downregulation on growth and pathogenesis of Cryptococcus neoformans[J]. Chinese Journal of Mycology, 2015, 10(1): 16-21.

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