Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2013 Jul 30;110(31):12750-5. doi: 10.1073/pnas.1310735110. Epub 2013 Jul 12.

RNA interference knockdown of DNA methyl-transferase 3 affects gene alternative splicing in the honey bee.

Author information

1
Department of Entomology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Abstract

Studies of DNA methylation from fungi, plants, and animals indicate that gene body methylation is ancient and highly conserved in eukaryotic genomes, but its role has not been clearly defined. It has been postulated that regulation of alternative splicing of transcripts was an original function of DNA methylation, but a direct experimental test of the effect of methylation on alternative slicing at the whole genome level has never been performed. To do this, we developed a unique method to administer RNA interference (RNAi) in a high-throughput and noninvasive manner and then used it to knock down the expression of DNA methyl-transferase 3 (dnmt3), which is required for de novo DNA methylation. We chose the honey bee (Apis mellifera) for this test because it has recently emerged as an important model organism for studying the effects of DNA methylation on development and social behavior, and DNA methylation in honey bees is predominantly on gene bodies. Here we show that dnmt3 RNAi decreased global genomic methylation level as expected and in addition caused widespread and diverse changes in alternative splicing in fat tissue. Four different types of splicing events were affected by dnmt3 gene knockdown, and change in two types, exon skipping and intron retention, was directly related to decreased methylation. These results demonstrate that one function of gene body DNA methylation is to regulate alternative splicing.

KEYWORDS:

epigenetics; gene regulation; gene silencing; insect

PMID:
23852726
PMCID:
PMC3732956
DOI:
10.1073/pnas.1310735110
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center