The RNA helicase Ski2 in the fungal pathogen Cryptococcus neoformans highlights key roles in azoles resistance and stress tolerance
The RNA helicase Ski2 in the fungal pathogen Cryptococcus neoformans highlights key roles in azoles resistance and stress tolerance
复制标题
真菌病原体新型隐球菌中的 RNA 解旋酶 Ski2 强调了在唑类抗性和应激耐受性中的关键作用
DOI：
10.1093/mmy/myac083
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发表时间：
2022
期刊：
影响因子：
2.9
通讯作者：
Xudong Zhu
中科院分区：
文献类型：
--
作者：
Chenxi Li;Xiaoyu Ma;Lan Ma;Sihui Zhen;Yinan Na;Ping Zhang;Xudong Zhu
抽象的。酵母菌SKI(超级杀手)复合体最初是从感染M‘杀手’病毒的细胞中鉴定出来的。Ski2作为SKI复合体的核心，是一种胞质辅因子，也是RNA降解外体的调节因子。推测的RNA解旋酶Ski2从酵母到动物都是高度保守的，已经被证明在调节RNA监视、温度敏感性和在几种酵母中的生长中发挥关键作用，但在新生隐球菌中还没有。在这里，我们报告了在真菌病原菌新生葡萄球菌中发现了一个编码等同的Ski2蛋白的基因，命名为Ski2。为了深入了解Ski2的功能，我们用CRISPR-Cas9编辑工具创建了一个突变株Ski2Δ。Ski2的破坏损害了细胞壁的完整性。进一步的研究发现，Ski2Δ突变体在抵抗渗透胁迫和极端生长温度方面存在缺陷。然而，值得注意的是，在营养耗竭的条件下，Ski2Δ突变体经历侵入性生长的能力增强了。更重要的是，我们的结果表明，与野生型相比，Ski2Δ突变体的毒力略低，对抗核糖体药物严重敏感，但它对唑类和氟胞嘧啶产生了多重耐药性。通过构建双缺失株Ski2Δafr1Δ，我们证实了Ski2Δ中Afr1基因的增加导致了对唑类的耐药性，这可能是非经典的小干扰RNA影响的结果。我们的工作表明Ski2在酵母病原菌新生葡萄球菌的耐药性和基因转录调控中起关键作用。
Abstract. The yeast SKI (superkiller) complex was originally identified from cells that were infected by the M ‘killer’ virus. Ski2, as the core of the SKI complex, is a cytoplasmic cofactor and regulator of RNA-degrading exosome. The putative RNA helicase Ski2 was highly conserved from yeast to animals and has been demonstrated to play a key role in the regulation of RNA surveillance, temperature sensitivity, and growth in several yeasts but not yet in Cryptococcus neoformans (C. neoformans). Here, we report the identification of a gene encoding an equivalent Ski2 protein, named SKI2, in the fungal pathogen C. neoformans. To obtain insights into the function of Ski2, we created a mutant strain, ski2Δ, with the CRISPR–Cas9 editing tool. Disruption of SKI2 impaired cell wall integrity. Further investigations revealed the defects of the ski2Δ mutant in resistance to osmotic stresses and extreme growth temperatures. However, significantly, the ability to undergo invasive growth under nutrient-depleted conditions was increased in the ski2Δ mutant. More importantly, our results showed that the ski2Δ mutant exhibited slightly lower virulence and severe susceptibility to anti-ribosomal drugs by comparison to the wild type, but it developed multidrug resistance to azoles and flucytosine. By constructing the double deletion strain ski2Δafr1Δ, we verified that increased Afr1 in ski2Δ contributed to the azole resistance, which might be influenced by nonclassical small interfering RNA. Our work suggests that Ski2 plays critical roles in drug resistance and regulation of gene transcription in the yeast pathogen C. neoformans.