| Congresso Brasileiro de Microbiologia 2023 | Resumo: 620-1 | ||||
Resumo:The regulation of gene expression in eukaryotes is knowingly responsive to stimuli through a network of competing signaling pathways. This constitutes a central system associated with transistor functions, tuning physiological responses and transcription rates to environmental conditions. Using a meta-transcriptomic workflow, we searched to determine the extent of downstream regulatory networks by alternative splicing (AS) for the control of gene expression in fungi. Our results support the occurrence of AS in a large subset of genes in RNA-Seq data with different viewpoints of the biology of the dermatophyte model species Trichophyton rubrum: growth in glucose- or keratin-supplemented medium (1,340 genes), exposure to the antifungal Undecanoic Acid (UDA) (654 genes), and knock-out mutations in the transcription factors (TF) genes ap1 (1,432 genes) and stuA (1,940 genes). In the set of 17 libraries, 5,449 AS events by Intron Retention (IR) were detected in 2,763 genes. Notably, this number represents 31.71% of the 8,713 genes found in the genome of T. rubrum. Most importantly, we highlight AS as a stress-responsive system. In fact, the number of splicing-regulated was equivalent to differentially expressed genes in two experiments: ap1 mutation and UDA exposure. Furthermore, we ran an analysis to search for AS specific to TF. Libraries from all experiments were enriched with IR events in TF. A total of 123 events were detected in 67 TF genes. The results revealed a clear overlap between splicing factors and TF responses. Enrichment Analysis and Gene Ontology (GO) categorization of AS events showed that libraries were also enriched with regulatory functions such as: translation, regulation of transcription, RNA binding, and mRNA processing. Additionally, we searched for the occurrence of proteins harboring prion domains (PrDs) in the genome of T. rubrum using Hidden Markov model predictions. Proteins performing diverse GO molecular functions were characterized in our list of 275 bona fide prion candidates, and we confirmed the current understanding that PrDs are mostly related to DNA and RNA-binding molecular functions. Interestingly, all 275 prion candidates were modulated in our experiments, either by AS or differential expression. Fifteen of these prion candidates were identified as TF. Altogether, these results show that compensatory mechanisms involving the transcription and splicing machinery must act concertedly. We showed that overlaps between these systems play an adaptive function during fungal metabolic reprogramming, providing fine-tuning of gene expression in response to stimuli. We have also determined that the environmental stress response program is enriched with PrD-encoding transcripts. PrDs have the innate tendency to form amyloid aggregates, and these transcripts are also modulated by splicing during stress. Palavras-chave: Alternative Splicing, Prions, Stress Response, Proteostasis, Transcriptome Agência de fomento:FAPESP | |||||