Identification of key structural determinants of the IntI1 integron integrase that influence attC x attI1 recombination efficiency
| Autor: | Didier Mazel, Deshmukh N. Gopaul, Gaëlle Demarre, Clara Frumerie |
|---|---|
| Přispěvatelé: | Plasticité du Génome Bactérien (PGB), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Biochimie et Biophysique des Macromolécules, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Models Molecular Mutant Molecular Sequence Data Mutagenesis (molecular biology technique) Electrophoretic Mobility Shift Assay Integron 03 medical and health sciences Genetics Recombinase [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Binding site 030304 developmental biology Gene Library Recombination Genetic 0303 health sciences Binding Sites biology Base Sequence Integrases 030306 microbiology Nucleic Acid Enzymes Integrase Mutagenesis Attachment Sites Microbiological biology.protein Nucleic Acid Conformation Recombination |
| Zdroj: | Nucleic Acids Research Nucleic Acids Research, Oxford University Press, 2007, epub ahead of print. ⟨10.1093/nar/gkm709⟩ Nucleic Acids Research, 2007, epub ahead of print. ⟨10.1093/nar/gkm709⟩ |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkm709⟩ |
| Popis: | The integron platform codes for an integrase (IntI) from the tyrosine family of recombinases that mediates recombination between a proximal double-strand recombination site, attI and a single-strand target recombination site, attC. The attI site is only recognized by its cognate integrase, while the various tested attCs sites are recombined by several different IntI integrases. We have developed a genetic system to enrich and select mutants of IntI1 that provide a higher yield of recombination in order to identify key protein structural elements important for attC x attI1 recombination. We isolated mutants with higher activity on wild type and mutant attC sites. Interestingly, three out of four characterized IntI1 mutants selected on different substrates are mutants of the conserved aspartic acid in position 161. The IntI1 model we made based on the VchIntIA 3D structure suggests that substitution at this position, which plays a central role in multimer assembly, can increase or decrease the stability of the complex and accordingly influence the rate of attI x attC recombination versus attC x attC. These results suggest that there is a balance between the specificity of the protein and the protein/protein interactions in the recombination synapse. |
| Databáze: | OpenAIRE |
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