Analysis of mitochondrial DNA copy number and its regulation through DNA methylation of POLGA

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)

Abstract

Replication of mitochondrial DNA (mtDNA) is important for ensuring that cells have sufficient mtDNA copy number to meet their specific requirements for the generation of cellular energy through oxidative phosphorylation. A number of transcription and replication factors are required for this process, with a key factor being the nuclear-encoded mtDNA-specific DNA polymerase γ. DNA polymerase γ has a catalytic subunit (POLGA), whose gene has been shown to be DNA methylated at exon 2. This methylation is considered to be one of the key mechanisms that regulate mtDNA copy number. These findings have made it of great importance to establish optimal methods for investigating the effects of DNA methylation on mtDNA replication. Here, we provide methods to determine the extent of DNA methylation at exon 2 of POLGA as well as other gene targets of interest. We also show how mtDNA copy number is assessed and, from these two outputs, define the efficiency of mtDNA replication by calculating the mtDNA-replicative efficiency index.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsMatthew McKenzie
PublisherHumana Press
Pages131-141
Number of pages11
Volume1351
Edition3rd
ISBN (Electronic)978-1-4939-3040-1
ISBN (Print)978-1-4939-3039-5
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1351
ISSN (Print)10643745

Keywords

  • DNA methylation
  • MeDIP
  • MtDNA copy number
  • POLGA

Cite this

Sun, X., Lee, W., Vaghjiani, V., & St. John, J. C. (2016). Analysis of mitochondrial DNA copy number and its regulation through DNA methylation of POLGA. In M. McKenzie (Ed.), Methods in Molecular Biology (3rd ed., Vol. 1351, pp. 131-141). (Methods in Molecular Biology; Vol. 1351). Humana Press. https://doi.org/10.1007/978-1-4939-3040-1_10