New function for the RNA helicase p68/DDX5 as a modifier of MBNL1 activity on expanded CUG repeats

Myotonic Dystrophy type I (DM1) is caused by an abnormal expansion of CTG triplets in the 3′ UTR of the dystrophia myotonica protein kinase (DMPK) gene, leading to the aggregation of the mutant transcript in nuclear RNA foci. The expanded mutant transcript promotes the sequestration of the MBNL1 splicing factor, resulting in the misregulation of a subset of alternative splicing events. In this study, we identify the DEAD-box RNA helicase p68 (DDX5) in complexes assembled onto in vitro-transcribed CUG repeats. We showed that p68 colocalized with RNA foci in cells expressing the 3′UTR of the DMPK gene containing expanded CTG repeats. We found that p68 increased MBNL1 binding onto pathological repeats and the stem–loop structure regulatory element within the cardiac Troponin T (TNNT2) pre-mRNA, splicing of which is misregulated in DM1. Mutations in the helicase core of p68 prevented both the stimulatory effect of the protein on MBNL1 binding and the colocalization of p68 with CUG repeats, suggesting that remodeling of RNA secondary structure by p68 facilitates MBNL1 binding. We also found that the competence of p68 for regulating TNNT2 exon 5 inclusion depended on the integrity of MBNL1 binding sites. We propose that p68 acts as a modifier of MBNL1 activity on splicing targets and pathogenic RNA

Handbau und Psychose

Gecorrigeerd via dispenserDiss. Freiburg I. Br.OPLADEN-RUG0

Diversity of wild and cultivated pearl millet accessions (Pennisetum glaucum [L.] R. Br.) in Niger assessed by microsatellite markers

Genetic diversity of crop species in sub-Sahelian Africa is still poorly documented. Among such crops, pearl millet is one of the most important staple species. In Niger, pearl millet covers more than 65% of the total cultivated area. Analyzing pearl millet genetic diversity, its origin and its dynamics is important for in situ and ex situ germplasm conservation and to increase knowledge useful for breeding programs. We developed new genetic markers and a high-throughput technique for the genetic analysis of pearl millet. Using 25 microsatellite markers, we analyzed genetic diversity in 46 wild and 421 cultivated accessions of pearl millet in Niger. We showed a significantly lower number of alleles and lower gene diversity in cultivated pearl millet accessions than in wild accessions. This result contrasts with a previous study using iso-enzyme markers showing similar genetic diversity between cultivated and wild pearl millet populations. We found a strong differentiation between the cultivated and wild groups in Niger. Analyses of introgressions between cultivated and wild accessions showed modest but statistically supported evidence of introgressions. Wild accessions in the central region of Niger showed introgressions of cultivated alleles. Accessions of cultivated pearl millet showed introgressions of wild alleles in the western, central, and eastern parts of Niger

Spatio-temporal dynamics of genetic diversity in Sorghum bicolor in Niger

The dynamics of crop genetic diversity need to be assessed to draw up monitoring and conservation priorities. However, few surveys have been conducted in centres of diversity. Sub-Saharan Africa is the centre of origin of sorghum. Most Sahel countries have been faced with major human, environmental and social changes in recent decades, which are suspected to cause genetic erosion. Sorghum is the second staple cereal in Niger, a centre of diversity for this crop. Niger was submitted to recurrent drought period and to major social changes during these last decades. We report here on a spatio-temporal analysis of sorghum genetic diversity, conducted in 71 villages covering the rainfall gradient and range of agro-ecological conditions in Niger's agricultural areas. We used 28 microsatellite markers and applied spatial and genetic clustering methods to investigate change in genetic diversity over a 26-year period (1976-2003). Global genetic differentiation between the two collections was very low (F (st) = 0.0025). Most of the spatial clusters presented no major differentiation, as measured by F (st), and showed stability or an increase in allelic richness, except for two of them located in eastern Niger. The genetic clusters identified by Bayesian analysis did not show a major change between the two collections in the distribution of accessions between them or in their spatial location. These results suggest that farmers' management has globally preserved sorghum genetic diversity in Niger

Analyse multiéchelle de la diversité génétique des sorghos : compréhension des processus évolutifs pour la conservation in situ = Multi-scale analysis of sorghum genetic diversity : understanding the evolutionary processes for in situ conservation

Using microsatellite genetic markers, we analyzed 1,518 sorghum samples collected at different spatial scales in Burkina Faso, Mali, Niger, and in the village of Wante, from the landrace to the country scale. Genetic diversity and differentiation parameters were estimated to assess the effects of the main evolutionary processes on sorghum genetic diversity. The genetic variability found within a variety is mainly the result of the sorghum reproduction biology and the genetic drift process caused by the limited number of reproductive individuals at the time of variety introduction into a household or each year when farmers select their seeds. At the village scale, a low correlation is observed between the diversity of vernacular names and the genetic diversity assessed by microsatellites. No spatial genetic structuration is observed among villages more than 30 kilometers apart. This point highlightss that traditional seed systems operate at a very local scale. in Mali, a similar proportion of allelic richness is observed along a longitudinal transect and a latitudinal gradient that crosses a larger range of agroclimatic conditions. At the country scale, sorghum exhibits more genetic diversity in Niger than in Mali despite a lower agroclimatic range in Niger. These results demonstrate that the diversity of human groups acts together with the agro-ecological factors to shape the structure of sorghum genetic diversity. An important proportion of the overall genetic diversity present in the Cirad (Centre de cooperation internationale en recherche agronomique pour le developpement) "core collection" is found in Mali and Niger. The tremendous diversity cultivated by farmers in traditional agroecosystems of Western Africa supports the relevance of in situ approaches for sorghum conservation programs in this region. Both conservation and crop improvement requirements can be achieved through a better use of local germplasm in decentralized breeding programs