TRIM33 switches off Ifnb1 gene transcription during the late phase of macrophage activation

Despite its importance during viral or bacterial infections, transcriptional regulation of the interferon-β gene (Ifnb1) in activated macrophages is only partially understood. Here we report that TRIM33 deficiency results in high, sustained expression of Ifnb1 at late stages of toll-like receptor-mediated activation in macrophages but not in fibroblasts. In macrophages, TRIM33 is recruited by PU.1 to a conserved region, the Ifnb1 Control Element (ICE), located 15 kb upstream of the Ifnb1 transcription start site. ICE constitutively interacts with Ifnb1 through a TRIM33-independent chromatin loop. At late phases of lipopolysaccharide activation of macrophages, TRIM33 is bound to ICE, regulates Ifnb1 enhanceosome loading, controls Ifnb1 chromatin structure and represses Ifnb1 gene transcription by preventing recruitment of CBP/p300. These results characterize a previously unknown mechanism of macrophage-specific regulation of Ifnb1 transcription whereby TRIM33 is critical for Ifnb1 gene transcription shutdown

Etude fonctionnelle de la voie micro-ARN dans la biologie des cellules tumorales

Les micro-ARNs (miRNAs) sont des ARNs de 20-22 nucléotides, transcrits à partir du génome, dont la fonction est de réguler l expression génique en s appariant à des ARNm cibles, inhibant ainsi leur traduction et/ou entrainant leur dégradation. Dans les cancers, l expression des miRNAs est fortement dérégulée. Une majorité de miRNAs est diminuée dans les tissus tumoraux par rapport aux tissus normaux, et un lien causal a été décrit entre inhibition globale des miRNAs et tumorigenèse. Par ailleurs, des miRNAs agissant comme des suppresseurs de tumeurs et d autres comme des oncogènes ont été décrits. Dans ce contexte impliquant de plus en plus les miRNAs dans les pathologies néoplasiques, l objectif de ce travail était d étudier le rôle de la voie miRNA dans la biologie des cellules tumorales. Afin d identifier des cellules tumorales dépendant de miRNAs oncogènes endogènes pour survivre ou proliférer, nous avons développé une stratégie d inhibition globale de la biogenèse des miRNAs en ciblant Drosha ou DGCR8, les deux composants du microprocesseur, complexe nucléaire de maturation des miRNAs. Cette stratégie nous a permis d identifier des lignées cellulaires tumorales dans lesquelles l inhibition du microprocesseur conduit à un phénotype d arrêt de prolifération durable. Nous avons mis à profit cette dépendance à la voie miRNA pour réaliser un crible positif de complémentation du défaut de prolifération observé grâce à l expression de miRNAs individuels. Nous avons ainsi pu mettre en évidence des miRNAs capables de soutenir individuellement la prolifération de ces cellules tumorales. Cette stratégie nous a également permis de montrer des différences fonctionnelles entre miRNAs homologues ou de la même famille. La recherche des cibles régulées par ces miRNAs nous a permis d élaborer des hypothèses concernant les cibles potentiellement impliquées dans le phénotype observé. Nous avons ainsi démontré la participation du suppresseur de tumeur PTEN à l arrêt de prolifération induit par l inhibition du microprocesseur. La stratégie d inhibition globale de la voie miRNA suivie d une complémentation phénotypique par des miRNAs individuels permet de s affranchir de la grande redondance de séquence et de fonction des miRNAs et devrait pouvoir s appliquer d une manière plus générale à l étude d autres processus régulés par les miRNAs.MicroRNAs (miRNAs) are 20-22 nucleotides RNAs, transcribed from the genome, which regulate gene expression by base-pairing to target mRNAs, thus inhibiting their translation and/or leading to their degradation. In cancers, miRNAs expression is strongly deregulated. A majority of miRNAs is diminished in tumoral tissues compared to normal tissues, and a causal link has been established between global inhibition of the miRNA pathway and tumorigenesis. In addition, miRNAs acting like tumor suppressors or oncogenes have been described. In this context of growing evidences implicating miRNAs in neoplasic diseases, this work aimed to investigate the role played by miRNA pathway in the biology of tumoral cells. In order to identify tumoral cells depending on endogenous oncogenic miRNAs to proliferate or survive, we developed a strategy of global inhibition of miRNAs biogenesis by targeting Drosha or DGCR8, the two components of the microprocessor , the nuclear miRNA maturation complex. This strategy allowed us to identify tumoral cell lines in which microprocessor inhibition led to a sustained growth arrest. We took advantage of this miRNA pathway dependency to screen for individual miRNAs able to complement the observed growth defect. This complementation screen allowed us to identify individual miRNAs able to sustain growth in those tumoral cells. This strategy also highlighted functional differences between homologous miRNAs or between miRNAs from the same family. The search for targets regulated by those miRNAs allowed us to develop hypothesis concerning the potential targets involved in the observed phenotype. By using this approach, we demonstrated that the tumor suppressor PTEN was involved in the growth arrest induced by microprocessor inhibition. The strategy of global miRNA pathway inhibition followed by phenotypic complementation by individual miRNAs allows overcoming the high sequence and function redundancy of miRNAs. We thus think it could be applied more generally to the study of other cellular processes regulated by miRNAs.PARIS11-SCD-Bib. électronique (914719901) / SudocSudocFranceF

Impairing the microRNA biogenesis pathway induces proteome modifications characterized by size bias and enrichment in antioxidant proteins

International audienc

A second nitrogen permease regulator in Saccharomyces cerevisiae

AbstractWe describe a Saccharomyces cerevisiae mutant affected in its urea and proline transport capacities, and a gene coding for a protein complementing this mutation. This protein is not membrane-embedded and contains two PEST sequences, often found in regulatory factors. The mRNA is not down-regulated under nitrogen catabolite repression, and is induced by urea and proline. In the mutant, the PUT4 mRNA encoding the proline permease is not affected, whereas the DUR3 mRNA, involved in urea active transport, is strongly increased. Our data suggest that this protein is a post-transcriptional regulator of nitrogen permeases

A genetic variant controls interferon-β gene expression in human myeloid cells by preventing C/EBP-β binding on a conserved enhancer

International audienceInterferon β (IFN-β) is a cytokine that induces a global antiviral proteome, and regulates the adaptive immune response to infections and tumors. Its effects strongly depend on its level and timing of expression. Therefore, the transcription of its coding gene IFNB1 is strictly controlled. We have previously shown that in mice, the TRIM33 protein restrains Ifnb1 transcription in activated myeloid cells through an upstream inhibitory sequence called ICE. Here, we show that the deregulation of Ifnb1 expression observed in murine Trim33-/- macrophages correlates with abnormal looping of both ICE and the Ifnb1 gene to a 100 kb downstream region overlapping the Ptplad2/Hacd4 gene. This region is a predicted myeloid super-enhancer in which we could characterize 3 myeloid-specific active enhancers, one of which (E5) increases the response of the Ifnb1 promoter to activation. In humans, the orthologous region contains several single nucleotide polymorphisms (SNPs) known to be associated with decreased expression of IFNB1 in activated monocytes, and loops to the IFNB1 gene. The strongest association is found for the rs12553564 SNP, located in the E5 orthologous region. The minor allele of rs12553564 disrupts a conserved C/EBP-β binding motif, prevents binding of C/EBP-β, and abolishes the activation-induced enhancer activity of E5. Altogether, these results establish a link between a genetic variant preventing binding of a transcription factor and a higher order phenotype, and suggest that the frequent minor allele (around 30% worldwide) might be associated with phenotypes regulated by IFN-β expression in myeloid cells

Periprocedural, early, and long-term risks of pacemaker implantation after atrioventricular nodal re-entry tachycardia ablation: a French nationwide cohort

International audienceAims Pacemaker implantation (PI) after atrioventricular nodal re-entry tachycardia (AVNRT) ablation is a dreadful complication. We aimed to assess periprocedural, early, and late risks for PI. Methods and results All 27 022 patients who underwent latest AVNRT ablation in France from 2009 to 2017, were identified in the nationwide medicalization database. A control group of 305 152 patients hospitalized for arm, leg, or skin injuries with no history of AVNRT or supraventricular tachycardia were selected. After propensity score matching, both groups had mean age of 53 +/- 18 years and were predominantly female (64%). During this 9-year period, 822 of 27 022 (3.0%) AVNRT patients underwent P1, with significant higher risk in propensity-matched AVNRT patients compared to propensity-matched controls [2.9% vs. 0.9%; hazard ratio 3.4 (2.9-3.9), P 6 months) risk periods. Annualized late risk per 100 AVNRT patients was 0.2%. In comparison to controls, excess risk was 0.2% in <30-year-old AVNRT patients; 0.7% in 30-50-year-old; 1.1% in 50-70-year-old and 6.5% over 70-year-olds. Risk for PI was also significantly different according to three procedural factors: centres, experience, and ablation date, with a 30% decrease since 2015. Conclusions Periprocedural, early, and late risks for PI were higher after AVNRT ablation compared to propensity-matched controls. Longer follow-up is needed as the excess risk seems to persist late after AVNRT ablation

TRIM33 switches off Ifnb1 gene transcription during the late phase of macrophage activation

International audienceDespite its importance during viral or bacterial infections, transcriptional regulation of the interferon-β gene (Ifnb1) in activated macrophages is only partially understood. Here we report that TRIM33 deficiency results in high, sustained expression of Ifnb1 at late stages of toll-like receptor-mediated activation in macrophages but not in fibroblasts. In macrophages, TRIM33 is recruited by PU.1 to a conserved region, the Ifnb1 Control Element (ICE), located 15 kb upstream of the Ifnb1 transcription start site. ICE constitutively interacts with Ifnb1 through a TRIM33-independent chromatin loop. At late phases of lipopolysaccharide activation of macrophages, TRIM33 is bound to ICE, regulates Ifnb1 enhanceosome loading, controls Ifnb1 chromatin structure and represses Ifnb1 gene transcription by preventing recruitment of CBP/p300. These results characterize a previously unknown mechanism of macrophage-specific regulation of Ifnb1 transcription whereby TRIM33 is critical for Ifnb1 gene transcription shutdown

Insights into the genetic structure of the cowpea pest Callosobruchus maculatus in Africa

The bean beetle Callosobruchus maculatus is a worldwide generalist legume seed pest, present in the tropics and subtropics. Despite its economic importance, little is known about the population structure and migration patterns of this bruchid beetle in its continent of origin, both at local and regional scales. Gaining knowledge in the demographic processes experienced by C. maculatus, in particular in its native area, might give clues to understand evolutionary forces that have driven its expansion in traditional agroecosystems. Here, we analyze the genetic structure of 23 African populations of C. maculatus located in eleven different countries, using nuclear (28S) and mitochondrial (CytB) markers, and further apply phylogenetic and demographic analyses. The 28S sequences showed low genetic variability. In contrast, the analysis of CytB indicated significant genetic differentiation between most populations, suggesting low levels of gene flow, even at a reduced spatial scale. The results obtained from the estimation of various demographic parameters suggest evidence of recent population expansion, in agreement with a scenario of long-distance dispersal through trade of legume seeds for human consumption and population growth related to its pest nature. Overall, our results show that biogeography, isolation processes and human-mediated dispersal events drive the genetic structure of this pest in Africa