Binding of human mitochondrial transcription factor A, an HMG box protein, to a four-way DNA junction

Biochem Biophys Res Commun. 2000 May 10;271(2):492-8. doi: 10.1006/bbrc.2000.2656.

Abstract

Mitochondrial transcription factor A (mtTFA), the only known transcription factor in mitochondria, is also implicated in maintenance of mitochondrial genome although little is elucidated about its molecular basis. mtTFA is a member of HMG box proteins family. Some HMG proteins bind with high affinity to four-way DNA junctions that mimic a Holliday structure, a putative intermediate in DNA recombination. To explore possible involvement of a Holliday-like structure in the maintenance of mitochondrial genome, we examine the binding of recombinant human mtTFA to a synthetic four-way DNA junction. The human mtTFA binds to the four-way DNA junction with an approximately 10-fold higher affinity than to the corresponding linear duplex DNA and with essentially the same affinity as to a 40-mer DNA containing the human mitochondrial light strand promoter sequence. The mtTFA binds to the four-way as a monomer. Both of the two HMG box domains of human mtTFA are required for the high affinity binding to the four-way junction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Mitochondrial / metabolism
  • DNA-Binding Proteins / metabolism*
  • Dose-Response Relationship, Drug
  • Gene Library
  • Glutathione Transferase / metabolism
  • HeLa Cells
  • High Mobility Group Proteins / metabolism*
  • Humans
  • Kinetics
  • Magnesium / metabolism
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Plasmids
  • Protein Binding
  • Recombinant Proteins / metabolism
  • Surface Plasmon Resonance
  • Time Factors
  • Trans-Activators / metabolism*
  • Xenopus Proteins*

Substances

  • DNA, Mitochondrial
  • DNA-Binding Proteins
  • High Mobility Group Proteins
  • Recombinant Proteins
  • Trans-Activators
  • XL-MTTFA protein, Xenopus
  • Xenopus Proteins
  • Glutathione Transferase
  • Magnesium