Function of the N-terminal segment of the RecA-dependent nuclease Ref
| Autor: | Michael M. Cox, Tayla M. Olsen, Angela J. Gruber, Rachel Dvorak |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Stereochemistry
Molecular Sequence Data Fluorescence Polarization Biology Cleavage (embryo) 03 medical and health sciences chemistry.chemical_compound Endonuclease Genetics Amino Acid Sequence Disulfides 030304 developmental biology chemistry.chemical_classification 0303 health sciences Nuclease Sequence Homology Amino Acid Nucleic Acid Enzymes 030302 biochemistry & molecular biology Amino acid Rec A Recombinases chemistry Biochemistry biology.protein Dimerization Cytokinesis Recombination DNA Cysteine |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 |
| Popis: | The bacteriophage P1 Ref (recombination enhancement function) protein is a RecA-dependent, HNH endonuclease. It can be directed to create targeted double-strand breaks within a displacement loop formed by RecA. The 76 amino acid N-terminal region of Ref is positively charged (25/76 amino acid residues) and inherently unstructured in solution. Our investigation of N-terminal truncation variants shows this region is required for DNA binding, contains a Cys involved in incidental dimerization and is necessary for efficient Ref-mediated DNA cleavage. Specifically, Ref N-terminal truncation variants lacking between 21 and 47 amino acids are more effective RecA-mediated targeting nucleases. We propose a more refined set of options for the Ref-mediated cleavage mechanism, featuring the N-terminal region as an anchor for at least one of the DNA strand cleavage events. |
| Databáze: | OpenAIRE |
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