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ABF1 binding sites in yeast RNA polymerase genes
F Della Seta1, I Treich, J M Buhler
1Département de Biologie, Centre d'Etudes Nucléaires de Saclay, Gif-sur-Yvette, France.
The Journal of Biological Chemistry
|September 5, 1990
Summary
Researchers identified a key protein, ABF1 (autonomous replication sequences and silencer-binding factor), that regulates yeast RNA polymerase genes. This factor binds specifically to the polymerase upstream box, aiding in gene expression control.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- Gene regulation is crucial for cellular function.
- Understanding transcription factor interactions is key to deciphering gene expression control.
Purpose of the Study:
- To identify and characterize trans-acting factors regulating yeast RNA polymerase genes RPC160 and RPC40.
- To elucidate the binding properties and consensus sequence of the identified factor.
Main Methods:
- Gel retardation assays
- DNase protection assays
- Mutational analysis
- Gel DNA-binding assay
Main Results:
- A single binding component interacts with the promoters of RPC160 and RPC40.
- This factor was identified as ABF1/SBF-B, known for binding to autonomous replication sequences and silencers.
- ABF1 binds the polymerase upstream box with high affinity (Kapp = 5.10(-10) M).
- Mutational analysis defined a nine-base pair binding site, leading to a consensus sequence (RTCRYB(N)4ACG).
Conclusions:
- ABF1 is a trans-acting factor regulating yeast RNA polymerase genes RPC160 and RPC40.
- The study provides a detailed characterization of ABF1 binding to the polymerase upstream box.
- The derived consensus sequence facilitates evaluation of ABF1 binding sites and related factors.