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Identification of protein complexes with quantitative proteomics in S. cerevisiae
Published on: March 4, 2009
Identification and purification of a protein that binds the yeast ARS consensus sequence
1Swiss Institute for Experimental Cancer Research, Lausanne, Switzerland.
Cell
|March 8, 1991
Summary
Researchers discovered a yeast protein, ARS consensus-binding protein (ACBP), that binds to DNA replication origins. This protein may play a crucial role in initiating DNA replication in yeast cells.
Area of Science:
- Molecular Biology
- Yeast Genetics
- DNA Replication
Background:
- DNA replication initiation is a fundamental process in cell biology.
- Specific DNA sequences, known as ARS elements, are recognized to initiate replication in yeast.
- The proteins involved in recognizing and binding to ARS elements are not fully characterized.
Purpose of the Study:
- To identify and characterize proteins that bind to the ARS consensus sequence in yeast.
- To elucidate the role of such proteins in the initiation of DNA replication.
Main Methods:
- Purification of the ARS consensus-binding protein (ACBP) using two-step chromatography.
- Analysis of ACBP binding to various ARS elements and their mutants.
- Characterization of ACBP as a single 67 kDa polypeptide.
Main Results:
- A 67 kDa yeast protein, ACBP, was purified to homogeneity.
- ACBP specifically binds to the ARS consensus sequence, with preferential binding to T-rich single strands.
- Mutations in the ARS consensus sequence significantly reduce ACBP binding affinity.
- ACBP recognizes multiple binding sites at the histone H4 ARS and ARS1, correlating with functional ARS boundaries.
Conclusions:
- The purified ARS consensus-binding protein (ACBP) is a key factor in recognizing ARS elements.
- ACBP's binding properties suggest its involvement in the early stages of DNA replication initiation.
- A model is proposed where ACBP facilitates the initiation of DNA replication at ARS sequences.
