The lncRNA RZE1 Controls Cryptococcal Morphological Transition.
The lncRNA RZE1 Controls Cryptococcal Morphological Transition.
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DOI：
10.1371/journal.pgen.1005692
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发表时间：
2015-11
期刊：
影响因子：
4.5
通讯作者：
Lin X
中科院分区：
文献类型：
--
作者：
Chacko N;Zhao Y;Yang E;Wang L;Cai JJ;Lin X
在真菌病原体新形隐球菌中，从酵母到菌丝的转换是性发育过程中减数分裂事件发生前的一个重要形态学过程。形态型也被认为与隐球菌的潜在毒力有关。先前的研究发现，调控因子Znf2是菌丝形成的关键决策者，也是一种抗毒因子。通过正向遗传筛选，我们发现长链非编码RNA (lncRNA) RZE1在ZNF2的上游调控酵母向菌丝的转化。我们发现RZE1主要在顺式中起作用，而在反式中作用较弱。有趣的是，RZE1的功能仅限于其原生细胞核。因此，RZE1似乎并不直接影响Znf2的翻译或Znf2蛋白的亚细胞定位。转录组分析表明，RZE1的缺失降低了ZNF2和ZNF2主要下游靶点的转录水平。此外，使用单分子荧光原位杂交（smFISH）的显微镜检查表明，RZE1的缺失增加了细胞核中ZNF2转录本的比例。综上所述，该lncRNA通过调节关键的形态发生调节因子Znf2来控制隐球菌从酵母到菌丝的转变。这是lncRNA在人类真菌病原体中的首次功能表征。考虑到隐球菌和其他真菌病原体基因组中潜在的大量lncRNAs，研究结果表明lncRNAs在真菌发育过程中作为额外的遗传调控层，可能有助于这些“简单”真核生物的复杂性。在高等真核生物的发育过程中，非蛋白调节因子的参与是最近才被发现的一个领域。非蛋白编码基因以前被称为基因组的暗物质，现在被认为在真核生物的生命中起着重要的调节作用。通过正向遗传筛选，我们鉴定出RZE1是调控隐球菌形态发生的关键lncRNA。我们进一步发现RZE1调控ZNF2的转录和转录输出，而ZNF2编码关键的形态发生转录因子。RZE1是人类真菌病原体中第一个功能明确的lncRNA。考虑到隐球菌和其他真菌病原体中潜在的大量lncrna， RZE1-ZNF2调控系统可以作为研究lncrna在真核病原体中的发育和毒力的范例。
In the fungal pathogen Cryptococcus neoformans, the switch from yeast to hypha is an important morphological process preceding the meiotic events during sexual development. Morphotype is also known to be associated with cryptococcal virulence potential. Previous studies identified the regulator Znf2 as a key decision maker for hypha formation and as an anti-virulence factor. By a forward genetic screen, we discovered that a long non-coding RNA (lncRNA) RZE1 functions upstream of ZNF2 in regulating yeast-to-hypha transition. We demonstrate that RZE1 functions primarily in cis and less effectively in trans. Interestingly, RZE1’s function is restricted to its native nucleus. Accordingly, RZE1 does not appear to directly affect Znf2 translation or the subcellular localization of Znf2 protein. Transcriptome analysis indicates that the loss of RZE1 reduces the transcript level of ZNF2 and Znf2’s prominent downstream targets. In addition, microscopic examination using single molecule fluorescent in situ hybridization (smFISH) indicates that the loss of RZE1 increases the ratio of ZNF2 transcripts in the nucleus versus those in the cytoplasm. Taken together, this lncRNA controls Cryptococcus yeast-to-hypha transition through regulating the key morphogenesis regulator Znf2. This is the first functional characterization of a lncRNA in a human fungal pathogen. Given the potential large number of lncRNAs in the genomes of Cryptococcus and other fungal pathogens, the findings implicate lncRNAs as an additional layer of genetic regulation during fungal development that may well contribute to the complexity in these “simple” eukaryotes. The involvement of non-protein regulators in developmental processes in higher eukaryotes is an area that has come to light more recently. Earlier known as the dark matter of the genome, the non-protein coding genes are now recognized for their important regulatory roles in the life of eukaryotes. Using forward genetic screen, we identified RZE1 as a lncRNA with key function in regulating morphogenesis in Cryptococcus. We further discovered that RZE1 regulates the transcription and transcript export of ZNF2, which encodes the key morphogenesis transcription factor. RZE1 is the first functionally characterized lncRNA in a human fungal pathogen. Given the potential large number of lncRNAs in Cryptococcus and other fungal pathogens, the RZE1-ZNF2 regulatory system could serve as a paradigm for the investigation of lncRNAs in development and virulence in eukaryotic pathogens.