Role of DNMT-1 in ova-induced mouse model of asthma

M. Verma, B.D. Chattopadhyay, B.N. Paul

Indian Institute of Toxicology Research, Lucknow, India

Asthma has long been recognized as a complex genetic disease mediated by exposures to a variety of environmental
triggers. Both genetic and environmental influences are important to asthma pathogenesis, some developed asthma
or wheezing illness as a result of occupational exposures, whereas others may become affected after exposure to
urban air pollution. Epigenetics is the study of heritable changes in gene expression that occur without directly
altering the DNA sequence. As such, epigenetic regulation provides an attractive mechanistic explanation, in
addition to gene-by-environment interactions, for some of the molecular events linking early exposures with later
disease (such as allergies, diabetes, neurodegenerative diseases, and cardiovascular diseases). One mechanism is
DNA methylation, the covalent addition of a methyl group to a cytosine residue in a CpG site (i.e., where a cytosine
lies next to guanine in the DNA sequence).This covalent addition of methyl group in the cytosine residues are
catalyzed by DNA methyltransferase enzyme-1(DNMT1). During the reporting period, we developed asthmatic
mice by a series of sensitization and challenge with an allergen, ovalbumin. Mice were found asthmatic based on
lung histopathology, lung function and Th2 cytokine profile. Using this mouse model of asthma, we studied
expression profile of DNMT1 by qRT2-PCR. We observed low expression of DNMT1 in lung, trachea and BALF
cells of asthmatic mice in comparison to normal. We further found that this low expression of DNMT1 in different
airways tissues was due to CpG methylation of DNMT1 promoter. CpG methylation was studied by methylation
specific PCR (MSP) and Bisulphite sequencing. CpG sites are generally clustered in high frequency near gene
promoters and these regions are referred to as CpG islands. The methylation states of CpG islands in turn may affect
gene activity and expression. Hence we conclude that low expression of DNMT1 in asthmapathogenesis is due to
methylation of the CpG island in the DNMT1 promoter.

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