Statistical Analysis of Readthrough Levels for Nonsense Mutations in Mammalian Cells Reveals a Major Determinant of Response to Gentamicin

The efficiency of translation termination depends on the nature of the stop codon and the surrounding nucleotides. Some molecules, such as aminoglycoside antibiotics (gentamicin), decrease termination efficiency and are currently being evaluated for diseases caused by premature termination codons. However, the readthrough response to treatment is highly variable and little is known about the rules governing readthrough level and response to aminoglycosides. In this study, we carried out in-depth statistical analysis on a very large set of nonsense mutations to decipher the elements of nucleotide context responsible for modulating readthrough levels and gentamicin response. We quantified readthrough for 66 sequences containing a stop codon, in the presence and absence of gentamicin, in cultured mammalian cells. We demonstrated that the efficiency of readthrough after treatment is determined by the complex interplay between the stop codon and a larger sequence context. There was a strong positive correlation between basal and induced readthrough levels, and a weak negative correlation between basal readthrough level and gentamicin response (i.e. the factor of increase from basal to induced readthrough levels). The identity of the stop codon did not affect the response to gentamicin treatment. In agreement with a previous report, we confirm that the presence of a cytosine in +4 position promotes higher basal and gentamicin-induced readthrough than other nucleotides. We highlight for the first time that the presence of a uracil residue immediately upstream from the stop codon is a major determinant of the response to gentamicin. Moreover, this effect was mediated by the nucleotide itself, rather than by the amino-acid or tRNA corresponding to the −1 codon. Finally, we point out that a uracil at this position associated with a cytosine at +4 results in an optimal gentamicin-induced readthrough, which is the therapeutically relevant variable

Readthrough of Premature Termination Codons in the Adenomatous Polyposis Coli Gene Restores Its Biological Activity in Human Cancer Cells

The APC tumor suppressor gene is frequently mutated in human colorectal cancer, with nonsense mutations accounting for 30% of all mutations in this gene. Reintroduction of the WT APC gene into cancer cells generally reduces tumorigenicity or induces apoptosis. In this study, we explored the possibility of using drugs to induce premature termination codon (PTC) readthrough (aminoglycosides, negamycin), as a means of reactivating endogenous APC. By quantifying the readthrough of 11 nonsense mutations in APC, we were able to identify those giving the highest levels of readthrough after treatment. For these mutations, we demonstrated that aminoglycoside or negamycin treatment led to a recovery of the biological activity of APC in cancer cell lines, and showed that the level of APC activity was proportional to the level of induced readthrough. These findings show that treatment with readthrough inducers should be considered as a potential strategy for treating cancers caused by nonsense mutations APC gene. They also provide a rational basis for identifying mutations responsive to readthrough inducers

Everyday vulnerabilities and ''social dispositions'' in the Malian Sahel, an indication for evaluating future adaptability to water crises?

International audienceSince the 1970s, precipitation in the Sahel has decreased and become very irregular, leading to widespread drought, whilst the human need for water has rapidly increased. A new ''dispositions''-based approach was adapted in order to analyse human interactions with environmental hazards and applied to the case of Hombori village in northeastern Mali. This article explores how the population and political stakeholders perceive, live with and respond to the increasing scarcity of water. It also explores how their current vulnerability and ability to cope with variations in available water resources indicate future adaptability to climate shocks. On the one hand, this research shows how the population copes with variations in water resource availability: the population's socio-spatial organisation explains the inhabitants' exposure to this problem and some of the factors affecting vulnerability, the elderly and women being the hardest hit. The water issue is generally managed on a ''day-to-day'' basis and considered a big problem only in the dry season, thus lowering any incentive for self-protection. The main two variables that could explain this kind of risk management are the conflicting local governance and current social rules. On the other hand, the discussion of results, based on a conceptual model of social responses, explains why these current ''social dispositions'' to cope with and even address the water scarcity issue do not guarantee future adaptability to climate change

Deleterious ABCA7 mutations and transcript rescue mechanisms in early onset Alzheimer’s disease

Abstract: Premature termination codon (PTC) mutations in the ATP-Binding Cassette, Sub-Family A, Member 7 gene (ABCA7) have recently been identified as intermediate-to-high penetrant risk factor for late-onset Alzheimer's disease (LOAD). High variability, however, is observed in downstream ABCA7 mRNA and protein expression, disease penetrance, and onset age, indicative of unknown modifying factors. Here, we investigated the prevalence and disease penetrance of ABCA7 PTC mutations in a large early onset AD (EOAD)-control cohort, and examined the effect on transcript level with comprehensive third-generation long-read sequencing. We characterized the ABCA7 coding sequence with next-generation sequencing in 928 EOAD patients and 980 matched control individuals. With MetaSKAT rare variant association analysis, we observed a fivefold enrichment (p = 0.0004) of PTC mutations in EOAD patients (3%) versus controls (0.6%). Ten novel PTC mutations were only observed in patients, and PTC mutation carriers in general had an increased familial AD load. In addition, we observed nominal risk reducing trends for three common coding variants. Seven PTC mutations were further analyzed using targeted long-read cDNA sequencing on an Oxford Nanopore MinION platform. PTC-containing transcripts for each investigated PTC mutation were observed at varying proportion (5-41% of the total read count), implying incomplete nonsense-mediated mRNA decay (NMD). Furthermore, we distinguished and phased several previously unknown alternative splicing events (up to 30% of transcripts). In conjunction with PTC mutations, several of these novel ABCA7 isoforms have the potential to rescue deleterious PTC effects. In conclusion, ABCA7 PTC mutations play a substantial role in EOAD, warranting genetic screening of ABCA7 in genetically unexplained patients. Long-read cDNA sequencing revealed both varying degrees of NMD and transcript-modifying events, which may influence ABCA7 dosage, disease severity, and may create opportunities for therapeutic interventions in AD

Agroforesterie et services écosystémiques en zone tropicale

Respectueux de l’environnement et garantissant une sécurité alimentaire soutenue par la diversification des productions et des revenus qu’ils procurent, les systèmes agroforestiers apparaissent comme un modèle prometteur d’agriculture durable dans les pays du Sud les plus vulnérables aux changements globaux. Cependant, ces systèmes agroforestiers ne peuvent être optimisés qu’à condition de mieux comprendre et de mieux maîtriser les facteurs de leurs productions. L’ouvrage présente un ensemble de connaissances récentes sur les mécanismes biophysiques et socio-économiques qui sous-tendent le fonctionnement et la dynamique des systèmes agroforestiers. Il concerne, d’une part les systèmes agroforestiers à base de cultures pérennes, telles que cacaoyers et caféiers, de régions tropicales humides en Amérique du Sud, en Afrique de l’Est et du Centre, d’autre part les parcs arborés et arbustifs à base de cultures vivrières, principalement de céréales, de la région semi-aride subsaharienne d’Afrique de l’Ouest. Il synthétise les dernières avancées acquises grâce à plusieurs projets associant le Cirad, l’IRD et leurs partenaires du Sud qui ont été conduits entre 2012 et 2016 dans ces régions. L’ensemble de ces projets s’articulent autour des dynamiques des systèmes agroforestiers et des compromis entre les services de production et les autres services socio-écosystémiques que ces systèmes fournissent