Transcription by RNA polymerase II (RNAPII) is a dynamic process with frequent variations in the elongation rate. However, the physiological relevance of variations in RNAPII elongation kinetics has remained unclear. Here we show in yeast that a RNAPII mutant that reduces the transcription elongation rate causes widespread changes in alternative polyadenylation (APA). We unveil two mechanisms by which APA affects gene expression in the slow mutant: 3 ′ UTR shortening and gene derepression by premature transcription termination of upstream interfering noncoding RNAs. Strikingly, the genes affected by these mechanisms are enriched for functions involved in phosphate uptake and purine synthesis, processes essential for maintenance of the intracellular nucleotide pool. As nucleotide concentration regulates transcription elongation, our findings argue that RNAPII is a sensor of nucleotide availability and that genes important for nucleotide pool maintenance have adopted regulatory mechanisms responsive to reduced rates of transcription elongation.

Prostatic steroid-binding protein (PBP) is the most abundant protein synthesized in the rat ventral prostate. The protein is under strict androgenic control and is made of two subunits containing the polypeptides C1, C2 and C3. Using an 35S-labelled cDNA probe, we have used quantitative in-situ hybridization to assess the regulation of polypeptide C1 mRNA levels by sex steroids in the adult male rat. Densitometric quantification of autoradiographic hybridization signals revealed that a significant decrease in C1 mRNA levels could be detected 5 h after castration. Levels of C1 mRNA decreased by 50% 2.5 days after castration, while undetectable levels were reached within 7 days. Administration of the potent androgen 5 alpha-dihydrotestosterone to castrated rats caused a progressive increase in C1 mRNA levels which became significant 5 h after the first injection, while prolonged treatment, for 3 and 7 days, caused 50 and 100% reversals respectively of the effect of castration on C1 mRNA levels. Similar results were obtained by dot-blot hybridization using the same 32P-labelled cDNA probe, thus confirming the specificity and quantification achieved by in-situ hybridization. Administration of oestradiol-17 beta to orchiectomized adult rats for 14 days had no effect on steady-state C1 mRNA levels. Progesterone, on the other hand, at the dose used (2 mg twice daily) caused a marked increase in C1 mRNA levels, measured by in-situ hybridization, which was completely reversed by concomitant administration of the pure antiandrogen flutamide. The present data clearly demonstrate that the expression of PBP C1 peptide mRNA is under strict androgenic control and is a very sensitive and specific parameter of androgenic activity. They also indicate that quantitative in-situ hybridization is a powerful, sensitive and most efficient tool to study the regulation of gene expression while, in addition, providing precise information about the site of mRNA localization as well as information about the histology of the tissue, particularly the heterogeneous nature of the acinar response to androgenic stimulation and deprivation.

Transcription by RNA polymerase I (RNAPI) represents most of the transcriptional activity in eukaryotic cells and is associated with the production of mature ribosomal RNA (rRNA). As several rRNA maturation steps are coupled to RNAPI transcription, the rate of RNAPI elongation directly influences processing of nascent pre-rRNA, and changes in RNAPI transcription rate can result in alternative rRNA processing pathways in response to growth conditions and stress. However, factors and mechanisms that control RNAPI progression by influencing transcription elongation rate remain poorly understood. We show here that the conserved fission yeast RNA-binding protein Seb1 associates with the RNAPI transcription machinery and promotes RNAPI pausing states along the rDNA. The overall faster progression of RNAPI at the rDNA in Seb1-deficient cells impaired cotranscriptional pre-rRNA processing and the production of mature rRNAs. Given that Seb1 also influences pre-mRNA processing by modulating RNAPII progression, our findings unveil Seb1 as a pause-promoting factor for RNA polymerases I and II to control cotranscriptional RNA processing.

Transcription termination of protein-coding genes in eukaryotic cells usually relies on a tight coordination between the cleavage and polyadenylation of the pre-mRNA, and 5′-3′ degradation of the downstream nascent transcript. Here we investigated the contribution of the essential fission yeast endonuclease Pac1, a homolog of human Drosha that cleaves hairpin RNA structures, in triggering polyadenylation-independent transcription termination. Using ChIP-sequencing in Pac1-deficient cells, we found that Pac1 triggers transcription termination at snRNA and snoRNA genes as well as at specific protein-coding genes. Notably, we found that Pac1-dependent premature termination occurred at two genes encoding conserved transmembrane transporters whose expression were strongly repressed by Pac1. Analysis by genome editing indicated that a stem-loop structure in the nascent transcript directs Pac1-mediated cleavage and that the regions upstream and downstream of the Pac1 cleavage site in the targeted mRNAs were stabilized by mutation of nuclear 3′-5′ and 5′-3′ exonucleases, respectively. Our findings unveil a premature transcription termination pathway that uncouples co-transcriptional RNA cleavage from polyadenylation, triggering rapid nuclear RNA degradation.

L’article analyse le processus d’institutionnalisation de l’euro numérique actuellement en projet au sein de la Banque centrale européenne. Pour ce faire, nous procédons à une analyse systématique des communications relatives à l’euro numérique publiées par la BCE et ses représentants. Cette analyse conduit à identifier les thèmes autour desquels l’institution communique de manière privilégiée et donc les éléments de légitimation qu’elle emploie. Nous montrons que la BCE opère un cadrage particulier du projet et de l’innovation monétaire qui doit en naître. Cette approche est ensuite comparée aux projets alternatifs portés par les acteurs de la société civile. Il apparaît que la BCE adopte une communication tendant à invisibiliser les implications sociales et politiques du projet d’euro numérique en l’abordant d’une manière principalement technique, ancrée dans des préoccupations économiques et notamment financières. De leur côté, les propositions alternatives ouvrent la voie à un euro numérique doté de finalités spécifiques (pouvant être diverses). Alors que la monnaie est, quelle que soit sa forme, d’abord une institution portée par la légitimité dont elle bénéficie et la confiance qui lui est accordée, aucun des projets formulés quant à l’euro numérique ne peut se prévaloir d’être le plus légitime. Nous discutons des implications pratiques et théoriques de cette situation pour la conduite du projet d’euro numérique ainsi que pour la gouvernance des banques centrales.