In vitro transcription of mitochondrial deoxyribonucleic acid from yeast

Georg Michaelis, Stephen Douglass, Ming Jer Tsai, Kim Burchiel, Richard S. Criddle

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Mitochondrial and nuclear DNAs from various wild-type and petite yeast strains have been highly purified by hydroxylapatite chromatography. Yeast mitochondrial, nuclear, and Escherichia coli DNA-dependent RNA polymerases were used to transcribe mitochondrial DNA of yeast into RNA. The RNA produced by these enzymes was used to study both the nature of the enzyme products and the properties of the template DNAs. The E. coli enzyme and mitochondrial enzyme transcribed the ribosomal genes of mitochondrial DNA as shown by competition with cold mitochondrial ribosomal RNA. The labeled in vitro synthesized RNAs were hybridized to various DNAs in an attempt to compare the extent of homology among these DNAs. Very little homology was indicated between nuclear and mitochondrial DNA of yeast. Total cell DNA from two petite mutants lacking the mitochondrial satellite band showed the same low degree of homology to mitochondrial DNA. Wild-type mitochondrial DNAs differed in their homologies to each other and appeared to contain different amounts of repetitive DNA. Mitochondrial DNAs from two petite mutants showed reduced homology to wild-type mitochondrial DNA. However, no additional sequences which differed from those of wild-type mitochondrial DNA could be detected in the mitochondrial DNA of one petite mutant studied, thus indicating a simple deletion mechanism for the origin of this strain. This petite mutant lacks 50-60% of the wild-type mitochondrial genome transcribed in vitro.

Original languageEnglish (US)
Pages (from-to)2026-2036
Number of pages11
JournalBiochemistry
Volume11
Issue number11
StatePublished - 1972
Externally publishedYes

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Transcription
Mitochondrial DNA
Yeast
Yeasts
DNA
RNA
Enzymes
Escherichia coli
Genes
In Vitro Techniques
Ribosomal RNA
Mitochondrial Genome
DNA-Directed RNA Polymerases
Durapatite
Chromatography
Satellites

ASJC Scopus subject areas

  • Biochemistry

Cite this

Michaelis, G., Douglass, S., Tsai, M. J., Burchiel, K., & Criddle, R. S. (1972). In vitro transcription of mitochondrial deoxyribonucleic acid from yeast. Biochemistry, 11(11), 2026-2036.

In vitro transcription of mitochondrial deoxyribonucleic acid from yeast. / Michaelis, Georg; Douglass, Stephen; Tsai, Ming Jer; Burchiel, Kim; Criddle, Richard S.

In: Biochemistry, Vol. 11, No. 11, 1972, p. 2026-2036.

Research output: Contribution to journalArticle

Michaelis, G, Douglass, S, Tsai, MJ, Burchiel, K & Criddle, RS 1972, 'In vitro transcription of mitochondrial deoxyribonucleic acid from yeast', Biochemistry, vol. 11, no. 11, pp. 2026-2036.
Michaelis G, Douglass S, Tsai MJ, Burchiel K, Criddle RS. In vitro transcription of mitochondrial deoxyribonucleic acid from yeast. Biochemistry. 1972;11(11):2026-2036.
Michaelis, Georg ; Douglass, Stephen ; Tsai, Ming Jer ; Burchiel, Kim ; Criddle, Richard S. / In vitro transcription of mitochondrial deoxyribonucleic acid from yeast. In: Biochemistry. 1972 ; Vol. 11, No. 11. pp. 2026-2036.
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