In Vitro Reconstitution of Human Mitochondrial Transcription

Methods Mol Biol. 2021:2192:35-41. doi: 10.1007/978-1-0716-0834-0_3.

Abstract

In vitro assay based on a reconstituted mitochondrial transcription system serves as a method of choice to probe the functional importance of proteins and their structural motifs. Here we describe protocols for transcription assays designed to probe activity of the human mitochondrial RNA polymerase and the transcription initiation complex using RNA-DNA scaffold and synthetic promoter templates.

Keywords: Mitochondrial transcription; POLRTM; Promoter; RNAP; TFAM; TFB2M.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA, Mitochondrial / chemistry
  • DNA, Mitochondrial / genetics
  • DNA-Binding Proteins / chemistry
  • DNA-Directed RNA Polymerases / chemistry*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • In Vitro Techniques
  • Methyltransferases / chemistry
  • Mitochondria / metabolism*
  • Mitochondrial Proteins / chemistry
  • Promoter Regions, Genetic
  • Protein Binding
  • Transcription Factors / chemistry
  • Transcription, Genetic*

Substances

  • DNA, Mitochondrial
  • DNA-Binding Proteins
  • Mitochondrial Proteins
  • TEFM protein, human
  • TFAM protein, human
  • Transcription Factors
  • Methyltransferases
  • TFB2M protein, human
  • DNA-Directed RNA Polymerases
  • POLRMT protein, human