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A temperature-sensitive lambda cI repressor functions on a modified operator in yeast cells by masking the TATA
Molecular & General Genetics : MGG
|August 30, 1995
Summary
Researchers modified the yeast TDH3 promoter with a phage lambda operator. This modification, along with cI repressor expression, reduced reporter gene transcription, demonstrating a novel gene regulation mechanism.
Area of Science:
- Molecular Biology
- Gene Regulation
- Yeast Genetics
Background:
- The TDH3 promoter is crucial for gene expression in yeast.
- TATA box elements are essential for transcription initiation.
- Bacteriophage lambda cI repressor protein regulates gene expression.
Purpose of the Study:
- To investigate the role of the TATA box in TDH3 promoter activity.
- To engineer a regulatable promoter using bacteriophage lambda repressor system.
- To explore novel mechanisms of transcriptional control in yeast.
Main Methods:
- Construction of TDH3 promoter derivatives with a phage lambda operator sequence.
- Replacement of the native TATA box with a synthetic operator containing a TATA site.
- Introduction of a temperature-sensitive cI repressor gene.
- Reporter gene assays to measure transcriptional activity.
Main Results:
- Reporter gene transcription was significantly reduced when cI repressor was expressed.
- Deletion of the native TDH3 TATA box yielded similar reductions in transcription.
- The cI repressor likely inhibits transcription by masking the TATA element.
- Temperature-sensitive cI repressor enabled temperature-dependent gene expression control.
Conclusions:
- The TATA box is essential for TDH3 promoter function.
- The phage lambda cI repressor can be used to control yeast gene expression.
- Repressor binding to operator sequences can effectively mask TATA elements, providing a tool for inducible gene expression.