The Integrator complex terminates promoter-proximal transcription at protein-coding genes, bioRxiv, 2019-08-06

SUMMARYThe transition of RNA polymerase II (Pol II) from initiation to productive elongation is a central, regulated step in metazoan gene expression. At many genes, Pol II pauses stably in early elongation, remaining engaged with the 25-60 nucleotide-long nascent RNA for many minutes while awaiting signals for release into the gene body. However, a number of genes display highly unstable promoter Pol II, suggesting that paused polymerase might dissociate from template DNA at these promoters and release a short, non-productive mRNA. Here, we report that paused Pol II can be actively destabilized by the Integrator complex. Specifically, Integrator utilizes its RNA endonuclease activity to cleave nascent RNA and drive termination of paused Pol II. These findings uncover a previously unappreciated mechanism of metazoan gene repression, akin to bacterial transcription attenuation, wherein promoter-proximal Pol II is prevented from entering productive elongation through factor-regulated termination.Highlights<jatslist list-type=bullet><jatslist-item>The Integrator complex inhibits transcription elongation at ∼15% of mRNA genes<jatslist-item><jatslist-item>Integrator targets promoter-proximally paused Pol II for termination<jatslist-item><jatslist-item>The RNA endonuclease of Integrator subunit 11 is critical for gene attenuation<jatslist-item><jatslist-item>Integrator-repressed genes are enriched in signaling and growth-responsive pathways<jatslist-item>

biorxiv genomics 100-200-users 2019

Assembly methods for nanopore-based metagenomic sequencing a comparative study, bioRxiv, 2019-08-02

ABSTRACTBackgroundMetagenomic sequencing has lead to the recovery of previously unexplored microbial genomes. In this sense, short-reads sequencing platforms often result in highly fragmented metagenomes, thus complicating downstream analyses. Third generation sequencing technologies, such as MinION, could lead to more contiguous assemblies due to their ability to generate long reads. Nevertheless, there is a lack of studies evaluating the suitability of the available assembly tools for this new type of data.FindingsWe benchmarked the ability of different short-reads and long-reads tools to assembly two different commercially available mock communities, and observed remarkable differences in the resulting assemblies depending on the software of choice. Short-reads metagenomic assemblers proved unsuitable for MinION data. Among the long-reads assemblers tested, Flye and Canu were the only ones performing well in all the datasets. These tools were able to retrieve complete individual genomes directly from the metagenome, and assembled a bacterial genome in only two contigs in the best scenario. Despite the intrinsic high error of long-reads technologies, Canu and Flye lead to high accurate assemblies (~99.4-99.8 % of accuracy). However, errors still had an impact on the prediction of biosynthetic gene clusters.ConclusionsMinION metagenomic sequencing data proved sufficient for assembling low-complex microbial communities, leading to the recovery of highly complete and contiguous individual genomes. This work is the first systematic evaluation of the performance of different assembly tools on MinION data, and may help other researchers willing to use this technology to choose the most appropriate software depending on their goals. Future work is still needed in order to assess the performance of Oxford Nanopore MinION data on more complex microbiomes.

biorxiv bioinformatics 100-200-users 2019

Genetic tool development in marine protists Emerging model organisms for experimental cell biology, bioRxiv, 2019-08-02

ABSTRACTMarine microbial eukaryotes underpin the largest food web on the planet and influence global biogeochemical cycles that maintain habitability. They are also remarkably diverse and provide insights into evolution, including the origins of complex life forms, as revealed through genome analyses. However, their genetic tractability has been limited to a few species that do not represent the broader diversity of eukaryotic life or some of the most environmentally relevant taxa. Here, we report on genetic systems developed as a community resource for experimental cell biology of aquatic protists from across the eukaryotic tree and primarily from marine habitats. We present evidence for foreign DNA delivery and expression in 14 species never before transformed, report on the advancement of genetic systems in 7 species, review of an already published transformation protocol in 1 species and discuss why the transformation of 17 additional species has not been achieved yet. For all protists studied in this community effort, we outline our methods, constructs, and genome-editing approaches in the context of published systems. The reported breakthroughs on genetic manipulation position the community to dissect cellular mechanisms from a breadth of protists, which will collectively provide insights into ancestral eukaryotic lifeforms, protein diversification and evolution of cellular pathways.

biorxiv ecology 100-200-users 2019

Negative selection on complex traits limits genetic risk prediction accuracy between populations, bioRxiv, 2019-08-02

Accurate genetic risk prediction is a key goal for medical genetics and great progress has been made toward identifying individuals with extreme risk across several traits and diseases (Collins and Varmus, 2015). However, many of these studies are done in predominantly European populations (Bustamante et al., 2011; Popejoy and Fullerton, 2016). Although GWAS effect sizes correlate across ancestries (Wojcik et al., 2019), risk scores show substantial reductions in accuracy when applied to non-European populations (Kim et al., 2018; Martin et al., 2019; Scutari et al., 2016). We use simulations to show that human demographic history and negative selection on complex traits result in population specific genetic architectures. For traits under moderate negative selection, ~50% of the heritability can be accounted for by variants in Europe that are absent from Africa. We show that this directly leads to poor performance in risk prediction when using variants discovered in Europe to predict risk in African populations, especially in the tails of the risk distribution. To evaluate the impact of this effect in genomic data, we built a Bayesian model to stratify heritability between European-specific and shared variants and applied it to 43 traits and diseases in the UK Biobank. Across these phenotypes, we find ~50% of the heritability comes from European-specific variants, setting an upper bound on the accuracy of genetic risk prediction in non-European populations using effect sizes discovered in European populations. We conclude that genetic association studies need to include more diverse populations to enable to utility of genetic risk prediction in all populations.

biorxiv genetics 100-200-users 2019

 

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