Direct sequencing of deleted mitochondrial DNA in myopathic patients

Biochem Biophys Res Commun. 1989 Oct 16;164(1):156-63. doi: 10.1016/0006-291x(89)91696-3.

Abstract

To investigate the mechanism of mitochondrial DNA deletion in human diseases, we amplified the deleted mitochondrial DNA of five patients with mitochondrial myopathy by using the polymerase chain reaction, and directly sequenced the crossover regions of the deleted mitochondrial DNA without cloning. In Patient 1, a 7-bp directly repeated sequence of 5'-ATCCCCA-3' was found at the boundaries of deleted segment spanning 7,039 bp between the ATPase 6 and the cytochrome b genes. In Patients 2, 3, and 4, a 13-bp sequence of 5'-ACCTCCCTCACCA-3' was found in the boundaries of deleted segment spanning 4,977 bp between the ATPase 8 and the ND5 genes. In Patient 5, a 3-bp sequence of 5'-CCT-3' was found in the boundaries of deleted segment spanning 3,717 bp between the ATPase 6 and the ND5 genes. Similar directly repeated sequences may contribute to mitochondrial DNA deletions in human degenerative diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Amino Acid Sequence
  • Autoradiography
  • Base Sequence
  • Chromosome Deletion
  • DNA, Mitochondrial / genetics*
  • DNA-Directed DNA Polymerase
  • Female
  • Humans
  • Male
  • Mitochondria, Muscle / metabolism*
  • Molecular Sequence Data
  • Muscular Diseases / genetics*
  • Polymerase Chain Reaction

Substances

  • DNA, Mitochondrial
  • DNA-Directed DNA Polymerase