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G-quadruplexes and gene expression in Arabidopsis thaliana

TLDR
A novel method for identifying G4s is introduced, which uses a machine learning approach trained on datasets derived from the high throughput sequencing of G4 structures, to study the prevalence of PG4s in the genome of Arabidopsis thaliana, the model plant.
Abstract
G-Quadruplexes (G4s) are four stranded DNA structures which form in regions with high GC content and high GC skew. Because of the dependence of G4 structure on specific sequences, it is possible to predict putative G4s (PG4s) throughout genomic sequence. PG4s are non-uniformly distributed in genomes, with higher densities within various genic features, particularly promoters, 5’ untranslated regions (UTRs) and coding sequences (CDSs). When they form G4s, these sequences can have a variety of implications for biological processes including replication, transcription, translation and splicing. Here, we introduce a novel method for identifying PG4s, which uses a machine learning approach trained on datasets derived from the high throughput sequencing of G4 structures. We apply this and other techniques, to study the prevalence of PG4s in the genome of Arabidopsis thaliana, the model plant. Finally, we study the effect of G4 stabilisation on gene expression in Arabidopsis, using the GQuadruplex binding agent N-methyl mesoporphyrin (NMM). We identify a family of genes which are strongly downregulated by NMM, and find that they contain large numbers of PG4s in their CDSs.

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References
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TL;DR: It is shown that ultrashort 4sU-tagging not only provides snapshot pictures of eukaryotic gene expression but also reveals global RNA processing kinetics at nucleotide resolution and provides the tools to study molecular mechanisms of RNA processing and their contribution to the regulation of gene expression.
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TL;DR: The results provide three important clues: common factor(s) exist and may shape both exons and introns; the ordinal reduction patterns may reflect a time-orderly evolution; and the larger first and last exons may be splicing-required.
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R-loop formation at Snord116 mediates topotecan inhibition of Ube3a-antisense and allele-specific chromatin decondensation

TL;DR: It is demonstrated that topotecan treatment stabilizes the formation of RNA:DNA hybrids (R loops) at G-skewed repeat elements within paternal Snord116, corresponding to increased chromatin decondensation and inhibition of Ube3a-ATS expression.
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Genome-wide analysis reveals regulatory role of G4 DNA in gene transcription

TL;DR: A model of G4 DNA-mediated stimulation of transcription with the hypothesis that PG4M(D500) contributes to gene transcription by maintaining the DNA in an open conformation, while the asymmetric distribution of PG4D500 considerably reduces the probability of blocking the progression of the RNA polymerase complex on the template strand.
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Unmasking alternative splicing inside protein-coding exons defines exitrons and their role in proteome plasticity.

TL;DR: It is demonstrated that at least some exitrons originate from ancestral coding exons, and a "splicing memory" hypothesis is proposed whereby upon intron loss imprints of former exon borders defined by vestigial splicing regulatory elements could drive the evolution of exitron splicing.