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Updated: Feb 20, 2026
08:12
High-throughput Screening for Protein-based Inheritance in S. cerevisiae
Published on: August 8, 2017
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The yeast 2-μm plasmid Raf protein contributes to plasmid inheritance by stabilizing the Rep1 and Rep2 partitioning
Mary E McQuaid1, Jordan B Pinder1, Niroshaathevi Arumuggam1
1Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia B3H 4R2, Canada.
Nucleic Acids Research
|October 20, 2017
Summary
The yeast 2-μm plasmid uses Rep proteins and Raf to ensure inheritance. Raf stabilizes Rep proteins, aiding plasmid partitioning and persistence, even when Rep interactions are impaired.
Area of Science:
- Molecular Biology
- Yeast Genetics
- Epigenetics
Background:
- The yeast 2-μm plasmid is a high-copy number genetic element crucial for host cell division.
- Efficient plasmid partitioning relies on Rep1, Rep2, and STB locus.
- Rep proteins and Raf regulate plasmid gene transcription.
Purpose of the Study:
- Identify domains and residues in Rep2 and Raf for protein association.
- Delineate the Rep2 DNA-binding domain.
- Understand the role of Raf in plasmid maintenance.
Main Methods:
- Protein interaction assays
- Sequence analyses
- Mutational approaches
Main Results:
- Rep2 and Raf interactions with Rep1 and Rep2 were characterized.
- Rep1 promotes STB association and stabilizes Rep protein levels.
- Rep2 mutants impaired in self-association or Rep1 association maintained inheritance with functional Raf.
Conclusions:
- Rep protein complexes are essential for partitioning and transcriptional repression.
- Raf stabilizes Rep partitioning proteins, contributing to 2-μm plasmid persistence.
- Raf's role extends beyond transcriptional regulation to plasmid stability.
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