Browsing by Subject "sequences"
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Item Open Access A computational screen for site selective A-to-I editing detects novel sites in neuron specific Hu proteins(2010) Ensterö, Mats; Akerborg, Orjan; Lundin, Daniel; Wang, Bei; Furey, Terrence S; Ohman, Marie; Lagergren, JensBackground: Several bioinformatic approaches have previously been used to find novel sites of ADAR mediated A-to-I RNA editing in human. These studies have discovered thousands of genes that are hyper-edited in their non-coding intronic regions, especially in alu retrotransposable elements, but very few substrates that are site-selectively edited in coding regions. Known RNA edited substrates suggest, however, that site selective A-to-I editing is particularly important for normal brain development in mammals. Results: We have compiled a screen that enables the identification of new sites of site-selective editing, primarily in coding sequences. To avoid hyper-edited repeat regions, we applied our screen to the alu-free mouse genome. Focusing on the mouse also facilitated better experimental verification. To identify candidate sites of RNA editing, we first performed an explorative screen based on RNA structure and genomic sequence conservation. We further evaluated the results of the explorative screen by determining which transcripts were enriched for A-G mismatches between the genomic template and the expressed sequence since the editing product, inosine (I), is read as guanosine (G) by the translational machinery. For expressed sequences, we only considered coding regions to focus entirely on re-coding events. Lastly, we refined the results from the explorative screen using a novel scoring scheme based on characteristics for known A-to-I edited sites. The extent of editing in the final candidate genes was verified using total RNA from mouse brain and 454 sequencing. Conclusions: Using this method, we identified and confirmed efficient editing at one site in the Gabra3 gene. Editing was also verified at several other novel sites within candidates predicted to be edited. Five of these sites are situated in genes coding for the neuron-specific RNA binding proteins HuB and HuD.Item Open Access Guyanagaster, a New Wood-Decaying Sequestrate Fungal Genus Related to Armillaria (Physalacriaceae, Agaricales, Basidiomycota)(2010) Henkel, TW; Smith, ME; Aime, MCPremise of the study : Sequestrate basidiomycete fungi (e. g. "gasteromycetes") have foregone ballistospory and evolved alternative, often elaborate mechanisms of basidiospore dispersal with highly altered basidioma morphology. Sequestrate fungi have independently evolved in numerous Agaricomycete lineages, confounding taxonomic arrangements of these fungi for decades. Understanding the multiple origins and taxonomic affinities of sequestrate fungi provides insight into the evolutionary forces that can drastically alter basidioma morphology. In the neotropical rainforests of the Guiana Shield, we encountered a remarkable sequestrate fungus fruiting directly on decaying hardwood roots. The fungus 'singular combination of traits include a wood-decaying habit; black, verrucose peridium; reduced stipe; and gelatinized basidiospore mass. Methods : Guyanagaster necrorhiza gen. et sp. nov. is described. Macro-and micromorphological characters were assessed and compared to most similar taxa. To determine the phylogenetic affinities of the fungus, DNA sequence data were obtained for the 18S, ITS, and 28S rDNA, RBP2, and EF1 alpha regions and subjected to single-and multi-gene analyses. DNA sequences from fungal vegetative organs growing on decaying woody roots confirmed the wood-inhabiting lifestyle of Guyanagaster. Key results : Guyanagaster is morphologically unique among sequestrate fungi worldwide. Phylogenetic evidence places Guyanagaster in close relation to the wood-decaying mushroom genus Armillaria in the Physalacriaceae (Agaricales, Agaricomycetes, Basidiomycota). Conclusions : Guyanagaster represents an independently evolved sequestrate form within the Physalacriaceae. Although molecular data confirm that Guyanagaster is closely related to Armillaria, the unusual features of this fungus suggest a case of radically divergent morphological evolution.