Mutations in the genes for mitochondrial RNA polymerase and a second mitochondrial transcription factor of Saccharomyces cerevisiae

1989 ◽  
Vol 219 (1-2) ◽  
pp. 125-128 ◽  
Author(s):  
Thomas Lisowsky ◽  
Georg Michaelis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcription Factor ◽  
Rna Polymerase ◽  
Mitochondrial Rna ◽  
Mitochondrial Transcription ◽  
Mitochondrial Transcription Factor ◽  
Mitochondrial Rna Polymerase

Purification and characterization of human mitochondrial transcription factor 1

1988 ◽  
Vol 8 (8) ◽  
pp. 3496-3509
Author(s):  
R P Fisher ◽  
D A Clayton
Keyword(s):  
Transcription Factor ◽  
Mitochondrial Dna ◽  
Dna Binding ◽  
Sodium Dodecyl Sulfate ◽  
Rna Polymerase ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Mitochondrial Transcription ◽  
Mitochondrial Transcription Factor ◽  
Transcription Factor 1

We purified to near homogeneity a transcription factor from human KB cell mitochondria. This factor, designated mitochondrial transcription factor 1 (mtTF1), is required for the in vitro recognition of both major promoters of human mitochondrial DNA by the homologous mitochondrial RNA polymerase. Furthermore, it has been shown to bind upstream regulatory elements of the two major promoters. After separation from RNA polymerase by phosphocellulose chromatography, mtTF1 was chromatographed on a MonoQ anion-exchange fast-performance liquid chromatography column. Analysis of mtTF1-containing fractions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single major polypeptide with an Mr of approximately 25,000. Centrifugation in analytical glycerol gradients indicated a sedimentation coefficient of approximately 2.5 S, consistent with a monomeric 25-kilodalton protein. Finally, when the 25-kilodalton polypeptide was excised from a stained sodium dodecyl sulfate-polyacrylamide gel and allowed to renature, it regained DNA-binding and transcriptional stimulatory activities at both promoters. Although mtTF1 is the only mitochondrial DNA-binding transcription factor to be purified and characterized, its properties, such as a high affinity for random DNA and a weak specificity for one of its target sequences, may typify this class of regulatory proteins.

scholarly journals Purification and characterization of human mitochondrial transcription factor 1.

1988 ◽  
Vol 8 (8) ◽  
pp. 3496-3509 ◽  
Author(s):  
R P Fisher ◽  
D A Clayton
Keyword(s):  
Transcription Factor ◽  
Mitochondrial Dna ◽  
Dna Binding ◽  
Sodium Dodecyl Sulfate ◽  
Rna Polymerase ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Mitochondrial Transcription ◽  
Mitochondrial Transcription Factor ◽  
Transcription Factor 1

We purified to near homogeneity a transcription factor from human KB cell mitochondria. This factor, designated mitochondrial transcription factor 1 (mtTF1), is required for the in vitro recognition of both major promoters of human mitochondrial DNA by the homologous mitochondrial RNA polymerase. Furthermore, it has been shown to bind upstream regulatory elements of the two major promoters. After separation from RNA polymerase by phosphocellulose chromatography, mtTF1 was chromatographed on a MonoQ anion-exchange fast-performance liquid chromatography column. Analysis of mtTF1-containing fractions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single major polypeptide with an Mr of approximately 25,000. Centrifugation in analytical glycerol gradients indicated a sedimentation coefficient of approximately 2.5 S, consistent with a monomeric 25-kilodalton protein. Finally, when the 25-kilodalton polypeptide was excised from a stained sodium dodecyl sulfate-polyacrylamide gel and allowed to renature, it regained DNA-binding and transcriptional stimulatory activities at both promoters. Although mtTF1 is the only mitochondrial DNA-binding transcription factor to be purified and characterized, its properties, such as a high affinity for random DNA and a weak specificity for one of its target sequences, may typify this class of regulatory proteins.

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