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Genetic and functional interactions between Mus81-Mms4 and Rad27
Min-Jung Kang1, Chul-Hwan Lee, Young-Hoon Kang
1Center for DNA Replication and Genome Instability, Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea.
Nucleic Acids Research
|July 28, 2010
Summary
The study reveals that Mus81-Mms4 endonuclease suppresses dna2 mutations by interacting with Rad27 (yeast Fen1). This interaction stimulates Rad27 activity, aiding DNA replication fork processing and resolving replication impediments.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Rad27 (yeast Fen1) and Dna2 are key endonucleases for Okazaki fragment processing in yeast.
- Mus81-Mms4 is a structure-specific endonuclease involved in resolving stalled replication forks and recombination intermediates.
Purpose of the Study:
- To investigate the functional and physical interaction between Mus81-Mms4 and Rad27.
- To elucidate the role of this interaction in suppressing dna2 mutational defects and its impact on DNA replication.
Main Methods:
- In vitro assays to assess the stimulation of Rad27 and Mus81-Mms4 activities.
- Analysis of protein-protein interactions, specifically involving the C-terminal fragment of Rad27.
- Genetic analysis of double mutants (rad27 mus81, rad27 mms4) to evaluate synergistic lethality.
Main Results:
- Mus81-Mms4 significantly stimulates Rad27 activity, restoring growth in dna2 mutants.
- Rad27 stimulates Mus81-Mms4 cleavage of various substrates, including regressed replication forks.
- Rad27's stimulation of Mus81-Mms4 is mediated by a protein-protein interaction, independent of Rad27's catalytic activity, requiring its C-terminal 64 amino acids.
Conclusions:
- Mus81-Mms4 and Rad27 collaborate in DNA replication to resolve structures that impede replication progression.
- The synergistic lethality of rad27 mus81 and rad27 mms4 double mutants underscores the critical importance of their combined function.
- The interaction network involving Rad27, Dna2, and Mus81-Mms4 is crucial for maintaining genomic stability during DNA replication.
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