Department of Genetics, Ribeirão Preto Medical School, São Paulo University, Ribeirão Preto, São Paulo, Brazil.
PLoS One. 2018 Apr 18;13(4):e0195871. doi: 10.1371/journal.pone.0195871. eCollection 2018.
Advances in the understanding of molecular systems depend on specific tools like the disruption of genes to produce strains with the desired characteristics. The disruption of any mutagen sensitive (mus) genes in the model fungus Neurospora crassa, i.e. mus-51, mus-52, or mus-53, orthologous to the human genes KU70, KU80, and LIG4, respectively, provides efficient tools for gene targeting. Accordingly, we used RNA-sequencing and reverse transcription-quantitative polymerase chain reaction amplification techniques to evaluate the effects of mus-52 deletion in N. crassa gene transcriptional modulation, and thus, infer its influence regarding metabolic response to extracellular availability of inorganic phosphate (Pi). Notably, the absence of MUS-52 affected the transcription of a vast number of genes, highlighting the expression of those coding for transcription factors, kinases, circadian clocks, oxi-reduction balance, and membrane- and nucleolus-related proteins. These findings may provide insights toward the KU molecular mechanisms, which have been related to telomere maintenance, apoptosis, DNA replication, and gene transcription regulation, as well as associated human conditions including immune system disorders, cancer, and aging.
对分子系统的认识的进展依赖于特定的工具，如基因的破坏，以产生具有所需特性的菌株。在模式真菌粗糙脉孢菌（Neurospora crassa）中，任何易突变（mus）基因的破坏，例如与人类基因 KU70、KU80 和 LIG4 分别同源的 mus-51、mus-52 或 mus-53，为基因靶向提供了有效的工具。因此，我们使用 RNA 测序和反转录定量聚合酶链反应扩增技术来评估 mus-52 在 N. crassa 基因转录调节中的缺失对基因转录的影响，从而推断其对细胞外无机磷酸盐（Pi）可用性的代谢反应的影响。值得注意的是，MUS-52 的缺失影响了大量基因的转录，突出了那些编码转录因子、激酶、昼夜节律钟、氧化还原平衡以及与膜和核仁相关的蛋白质的基因的表达。这些发现可能为 KU 分子机制提供了一些见解，这些机制与端粒维持、细胞凋亡、DNA 复制和基因转录调节有关，以及与免疫系统紊乱、癌症和衰老等相关的人类疾病有关。
