Intronic Non-CG DNA Hydroxymethylation and Alternative mRNA Splicing in Honey Bees
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ABSTRACT:
PROVIDER: PRJNA219484 | ENA |
REPOSITORIES: ENA
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Project description:Honey bee non-CG DNA hydroxymethylation is enriched in the introns, which supplements previous findings that honey bee CG DNA methylation is enriched in exons.
2013-09-19 | GSE50990 | GEO
Project description:Honey bee non-CG DNA hydroxymethylation is enriched in the introns, which supplements previous findings that honey bee CG DNA methylation is enriched in exons. Bisulfite sequencing combined with Pvu-Seq to distinguish 5-methylcytosine from 5-hydroxymethylcytosine and RNA-Seq
2013-09-19 | E-GEOD-50990 | biostudies-arrayexpress
Project description:Previous whole-genome shotgun bisulfite sequencing experiments showed that DNA cytosine methylation in the honey bee (Apis mellifera) is almost exclusively at CG dinucleotides in exons. However, the most commonly used method, bisulfite sequencing, cannot distinguish 5-methylcytosine from 5-hydroxymethylcytosine, an oxidized form of 5-methylcytosine that is catalyzed by the TET family of dioxygenases. Furthermore, some analysis software programs under-represent non-CG DNA methylation and hydryoxymethylation for a variety of reasons. Therefore, we used an unbiased analysis of bisulfite sequencing data combined with molecular and bioinformatics approaches to distinguish 5-methylcytosine from 5-hydroxymethylcytosine. By doing this, we have performed the first whole genome analyses of DNA modifications at non-CG sites in honey bees and correlated the effects of these DNA modifications on gene expression and alternative mRNA splicing.We confirmed, using unbiased analyses of whole-genome shotgun bisulfite sequencing (BS-seq) data, with both new data and published data, the previous finding that CG DNA methylation is enriched in exons in honey bees. However, we also found evidence that cytosine methylation and hydroxymethylation at non-CG sites is enriched in introns. Using antibodies against 5-hydroxmethylcytosine, we confirmed that DNA hydroxymethylation at non-CG sites is enriched in introns. Additionally, using a new technique, Pvu-seq (which employs the enzyme PvuRts1l to digest DNA at 5-hydroxymethylcytosine sites followed by next-generation DNA sequencing), we further confirmed that hydroxymethylation is enriched in introns at non-CG sites.Cytosine hydroxymethylation at non-CG sites might have more functional significance than previously appreciated, and in honey bees these modifications might be related to the regulation of alternative mRNA splicing by defining the locations of the introns.
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Project description:Background: Honey bee is a major insect used for pollination of a number of commercial crops worldwide. However, the number of managed honey bee colonies has recently declined in several countries, and a number of possible causes are proposed. Although the use of honey bees for pollination can be considered as disruption of the habitat, its effects on honey bees' physiology have never been addressed. In Japan, more than 100 thousands colonies are annually used for pollination, and intriguingly 80% of them are used in greenhouses. Recently, honey bee colonies have often collapsed when they are introduced into greenhouses. Thus, to suppress colony collapses and maintain the number of worker bees in the colonies are essential for successful long-term pollination in greenhouses and recycling honey bee colonies. Honey bee hives were installed into strawberry and eggplant greenhouses, and then the gene expression profiles of the honey bees were examined at the different time periods. Total 16 samples with two replicates were analyzed.
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Project description:The microsporidia Nosema ceranae are intracellular parasites that proliferate in the midgut epithelial cells of honey bees (Apis mellifera). To analyze the pathological effects of those microsporidia, we orally infected honey bee workers 7 days after their emergence. Bees were flash frozen 15 days after the infection. Then, the effects on the gut ventriculi were analyzed and compared to non-infected (control) bees. Comparisons of control vs Nosema ceranae bees
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