Mutations’ level detection of mitochondrial genome G14459A in homogenates of the human aortic intima

Abstract

Objective: The aim of this study was the pilot analysis of the heteroplasmy level for mutation G14459A of human
mitochondrial genome.
Materials and Methods: The DNA samples were isolated from total homogenates of normal and affected by
atherosclerosis aortic intima of individuals by a method of the phenol-chloroform extraction. After PCR the amplificates
were pyrosequenced to identify the percent of heteroplasmy.
Results and discussion: A single nucleotide substitution G14459A was analyzed. The mutation is localized in
the coding region of the mitochondrial genome, in particular, in the gene NADH dehydrogenase subunit 6. It causes
a defect of the protein subunit of mitochondrial respiratory chain enzyme, leading to dysfunction of the NADH
dehydrogenase and possibly to oxidative stress in the body.
It was found that the level of heteroplasmy for this mutation is significantly higher in total homogenates of affected by
atherosclerosis intima compared to homogenates of normal intima.
Conclusion: According to the data resulting from the study, G14459A mutation is associated with atherosclerotic
lesions of human aortic intima.