In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
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Ursula Oberholzer
Abstract
Genetic experiments in the yeast Saccharomyces cerevisiae have identified the five Not proteins as global repressors of transcription which preferentially repress core promoters which do not contain a canonical TATA sequence. Recently, the Ccr4 and Caf1 proteins, required for non-fermentative gene expression, were found to be associated with the five Not proteins in 1.2 and 2 MDa Ccr4-Not complexes. These Ccr4-Not complexes, as many other global regulators of transcription, appear to regulate transcription both positively and negatively in vivo.
To further characterize the activity of Not1p, the only essential known protein of the Ccr4-Not complex, and determine whether it can act directly as a transcriptional repressor, we established an in vitro transcription system in which the HIS3 TATA-less promoter can be efficiently transcribed. We demonstrate that transcription from the HIS3 TATA-less promoter can be specifically increased in vitro by preparing nuclear extracts from a conditional mutant of the NOT1 gene and analyzing transcription after shifting the nuclear extracts to the restrictive temperature. This result is the first demonstration that one of the Not proteins directly represses transcription. Moreover, it now defines an experimental system in which TATA-less transcription initiation and repression by the Ccr4-Not complex can be studied further.
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
Artikel in diesem Heft
- To Our Authors, Readers and Subscribers
- Editor's Note
- A Chimpanzee Millennium
- Molecular Genetics of Dopa-Responsive Dystonia
- In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
- Fast Control of DNA Replication in Response to Hypoxia and to Inhibited Protein Synthesis in CCRF-CEM and HeLa Cells
- The Inhibition of NF-B Activation Pathways and the Induction of Apoptosis by Dithiocarbamates in T Cells Are Blocked by the Glutathione Precursor N-Acetyl-L-Cysteine
- Xylose Utilisation: Cloning and Characterisation of the Xylose Reductase from Candida tenuis
- Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis
- Thermodynamic Properties and DNA Binding of the ParD Protein from the Broad Host-Range Plasmid RK2/RP4 Killing System
- Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
- Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
- Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
- The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
- Characterization of a New Member of the TNF Family Expressed on Antigen Presenting Cells
- Vascular Endothelial Growth Factor (VEGF) and Its Receptors in Tumor-Bearing Dogs
- Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
- Mitochondria-Derived and Extra-Mitochondrial Human Type-1 Porin Are Identical as Revealed by Amino Acid Sequencing and Electrophysiological Characterisation
- Acknowledgement
- Content Index
- Author Index
- Subject Index
Artikel in diesem Heft
- To Our Authors, Readers and Subscribers
- Editor's Note
- A Chimpanzee Millennium
- Molecular Genetics of Dopa-Responsive Dystonia
- In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
- Fast Control of DNA Replication in Response to Hypoxia and to Inhibited Protein Synthesis in CCRF-CEM and HeLa Cells
- The Inhibition of NF-B Activation Pathways and the Induction of Apoptosis by Dithiocarbamates in T Cells Are Blocked by the Glutathione Precursor N-Acetyl-L-Cysteine
- Xylose Utilisation: Cloning and Characterisation of the Xylose Reductase from Candida tenuis
- Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis
- Thermodynamic Properties and DNA Binding of the ParD Protein from the Broad Host-Range Plasmid RK2/RP4 Killing System
- Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
- Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
- Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
- The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
- Characterization of a New Member of the TNF Family Expressed on Antigen Presenting Cells
- Vascular Endothelial Growth Factor (VEGF) and Its Receptors in Tumor-Bearing Dogs
- Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
- Mitochondria-Derived and Extra-Mitochondrial Human Type-1 Porin Are Identical as Revealed by Amino Acid Sequencing and Electrophysiological Characterisation
- Acknowledgement
- Content Index
- Author Index
- Subject Index
