Changes in liver cell DNA methylation status in diabetic mice affect its FT-IR characteristics

PLoS One. 2014 Jul 14;9(7):e102295. doi: 10.1371/journal.pone.0102295. eCollection 2014.

Abstract

Background: Lower levels of cytosine methylation have been found in the liver cell DNA from non-obese diabetic (NOD) mice under hyperglycemic conditions. Because the Fourier transform-infrared (FT-IR) profiles of dry DNA samples are differently affected by DNA base composition, single-stranded form and histone binding, it is expected that the methylation status in the DNA could also affect its FT-IR profile.

Methodology/principal findings: The DNA FT-IR signatures obtained from the liver cell nuclei of hyperglycemic and normoglycemic NOD mice of the same age were compared. Dried DNA samples were examined in an IR microspectroscope equipped with an all-reflecting objective (ARO) and adequate software.

Conclusions/significance: Changes in DNA cytosine methylation levels induced by hyperglycemia in mouse liver cells produced changes in the respective DNA FT-IR profiles, revealing modifications to the vibrational intensities and frequencies of several chemical markers, including νas -CH3 stretching vibrations in the 5-methylcytosine methyl group. A smaller band area reflecting lower energy absorbed in the DNA was found in the hyperglycemic mice and assumed to be related to the lower levels of -CH3 groups. Other spectral differences were found at 1700-1500 cm(-1) and in the fingerprint region, and a slight change in the DNA conformation at the lower DNA methylation levels was suggested for the hyperglycemic mice. The changes that affect cytosine methylation levels certainly affect the DNA-protein interactions and, consequently, gene expression in liver cells from the hyperglycemic NOD mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytosine / metabolism
  • DNA / chemistry*
  • DNA / metabolism*
  • DNA Methylation*
  • Diabetes Mellitus, Experimental / genetics
  • Diabetes Mellitus, Experimental / metabolism*
  • Diabetes Mellitus, Experimental / pathology
  • Female
  • Liver / pathology*
  • Mice
  • Mice, Inbred NOD
  • Spectroscopy, Fourier Transform Infrared*

Substances

  • Cytosine
  • DNA

Grants and funding

This work was supported by grant no. 201050015-6 from the São Paulo State Research Foundation (FAPESP) and grants no. 301943/2009-5 and 475261/2012-7 from the Brazilian National Council for Research and Development (CNPq). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.