High-throughput transcriptome sequencing and comparative analysis of Escherichia coli and Schizosaccharomyces pombe in respiratory and fermentative growth.
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ABSTRACT:
SUBMITTER: Vichi J
PROVIDER: S-EPMC7968713 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Next Generation sequencing techniques have ushered in a new era in life sciences, especially genomic research. Currently, massive RNA sequencing (RNA-seq) has increased knowledge about the "RNA universe" from the global characterization of transcriptomes, differential gene expression studies and the continued discovery of non-coding regulatory transcripts such as non-coding RNAs (ncRNAs). Post-transcriptional gene regulation is a mechanism for controlling gene expression that is present in both prokaryote and eukaryote organisms and regulates multiple biological processes. Examples of ncRNAs that regulate the levels of transcripts of genes related to central carbon metabolism have been described in bacteria and higher eukaryotes. In order to describe whether analogies exist in regulatory circuits mediated by ncRNAs in two cell patterns, a comparative analysis of the transcriptome of Escherichia coli and Schizosaccharomyces pombe in similar growth conditions was carried out using two carbon source, 2 % glucose and a mix of 2% glycerol and 0.2 % sodium acetate. Twelve RNA-seq libraries were performed. The RNA-seq data contained a number unambiguously mapped readings on the genome of each organism ranged from 1-1.3 million. A total of 739 and 743 genes were detected as differentially expressed for E. coli and S, pombe respectively with a module of log FC > 1 and FDR > 0.05. In aerobic glicolisis GO asciated to cell proliferetionweres detected in both organisms. Among the genes differentially expressed in glycerol/acetate general transcription factors such as fliA, 11 literature validated sRNAs, and assembly flagellum maquinary gene were detected in E. coli, whereas in fission yeast, hexose transporters, switches integral membrane proteins and ncRNAs related to cellular stress are highlighted. In general, our study shows that a conserved "foraging behaviour" response is observed in eukaryotic and procaryotic organism when there is no present glycolityc carbon source.
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Project description:Eukaryotic telomeres are transcribed into telomeric repeat-containing RNA (TERRA). Telomeric transcription has been documented in mammals, birds, zebra fish, plants and budding yeast. Here we show that the chromosome ends of Schizosaccharomyces pombe produce distinct RNA species. As with budding yeast and mammals, S. pombe contains G-rich TERRA molecules and subtelomeric RNA species transcribed in the opposite direction of TERRA (ARRET). Moreover, fission yeast chromosome ends produce two novel RNA species: C-rich telomeric repeat-containing transcripts (ARIA) and subtelomeric transcripts complementary to ARRET (αARRET). RNA polymerase II (RNAPII) associates with pombe chromosome ends in vivo and the telomeric factor Rap1 negatively regulates this association, as well as the cellular accumulation of RNA emanating from chromosome ends. We also show that the RNAPII subunit Rpb7 and the non-canonical poly(A) polymerases Cid12 and Cid14 are involved in the regulation of TERRA, ARIA, ARRET and αARRET transcripts. We confirm the evolutionary conservation of telomere transcription, and reveal intriguing similarities and differences in the composition and regulation of telomeric transcripts among model organisms.
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Project description:The biological impact of transposons on the physiology of the host depends greatly on the frequency and position of integration. Previous studies of Tf1, a long terminal repeat retrotransposon in Schizosaccharomyces pombe, showed that integration occurs at the promoters of RNA polymerase II (Pol II) transcribed genes. To determine whether specific promoters are preferred targets of integration, we sequenced large numbers of insertions using high-throughput pyrosequencing. In four independent experiments we identified a total of 73,125 independent integration events. These data provided strong support for the conclusion that Pol II promoters are the targets of Tf1 integration. The size and number of the integration experiments resulted in reproducible measures of integration for each intergenic region and ORF in the S. pombe genome. The reproducibility of the integration activity from experiment to experiment demonstrates that we have saturated the full set of insertion sites that are actively targeted by Tf1. We found Tf1 integration was highly biased in favor of a specific set of Pol II promoters. The overwhelming majority (76%) of the insertions were distributed in intergenic sequences that contained 31% of the promoters of S. pombe. Interestingly, there was no correlation between the amount of integration at these promoters and their level of transcription. Instead, we found Tf1 had a strong preference for promoters that are induced by conditions of stress. This targeting of stress response genes coupled with the ability of Tf1 to regulate the expression of adjacent genes suggests Tf1 may improve the survival of S. pombe when cells are exposed to environmental stress.
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Project description:The fission yeast Schizosaccharomyces pombe is a widely used model organism to study basic mechanisms of eukaryotic biology, but unlike other model organisms, its proteome remains largely uncharacterized. Using a shotgun proteomics approach based on multidimensional prefractionation and tandem mass spectrometry, we have detected approximately 30% of the theoretical fission yeast proteome. Applying statistical modelling to normalize spectral counts to the number of predicted tryptic peptides, we have performed label-free quantification of 1465 proteins. The fission yeast protein data showed considerable correlations with mRNA levels and with the abundance of orthologous proteins in budding yeast. Functional pathway analysis indicated that the mRNA-protein correlation is strong for proteins involved in signalling and metabolic processes, but increasingly discordant for components of protein complexes, which clustered in groups with similar mRNA-protein ratios. Self-organizing map clustering of large-scale protein and mRNA data from fission and budding yeast revealed coordinate but not always concordant expression of components of functional pathways and protein complexes. This finding reaffirms at the protein level the considerable divergence in gene expression patterns of the two model organisms that was noticed in previous transcriptomic studies.
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