Abstract
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.
Highlights
Changes in the environment, including nutrient availability, can be sensed by all living organisms
Using the experimental setup described in the Material and Methods section with an adjusted P-value 0.05 and an independent filter [20], we identified a set of differentially expressed genes (DEGs) (S1 Table)
For the mutant FGSC 9568 strain cultured in the same Pi availability condition we have identified 144 DEGs, comprising 4 up- and 140 down-regulated genes; from these, 2 up- (NCU11307 and NCU00552) and 4 down-regulated (NCU03921, NCU04148, NCU09627, NCU00878) genes were highlighted as related to integral membrane components (Table B in S2 Table)
Summary
Changes in the environment, including nutrient availability, can be sensed by all living organisms. Living systems react to changes with appropriate cellular responses, such as regulation of growth, proliferation, metabolism, and apoptosis. The uptake and utilization of nutrients including inorganic phosphate (Pi) is crucial in physiological metabolisms, such as energy transduction, maintenance of genetic information, cell growth, synthesis of membrane phospholipids, and cellular signalling processes in general [4]. The genetic and molecular mechanisms controlling the response to Pi starvation in N. crassa include at least. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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