Tailoring the activity of restriction endonuclease Ple I by PNA‐induced DNA looping
| Autor: | Maxim D. Frank-Kamenetskii, Ekaterina Protozanova, Viatcheslav A. Soldatenkov, Sergey Chasovskikh, Vadim V. Demidov |
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| Rok vydání: | 2002 |
| Předmět: |
Time Factors
DNA clamp Base Sequence HMG-box Scientific Report Molecular Sequence Data DNA Sequence Analysis DNA Nicking enzyme Biology Microscopy Atomic Force Biochemistry Restriction fragment DNA binding site Restriction enzyme Restriction map Genetics biology.protein Nucleic Acid Conformation Deoxyribonucleases Type II Site-Specific Peptides Dimerization Molecular Biology In vitro recombination |
| Zdroj: | EMBO reports. 3:956-961 |
| ISSN: | 1469-3178 1469-221X |
| DOI: | 10.1093/embo-reports/kvf192 |
| Popis: | DNA looping is one of the key factors allowing proteins bound to different DNA sites to signal one another via direct contacts. We demonstrate that DNA looping can be generated in an arbitrary chosen site by sequence-directed targeting of double-stranded DNA with pseudocomplementary peptide-nucleic acids (pcPNAs). We designed pcPNAs to mask the DNA from cleavage by type IIs restriction enzyme PleI while not preventing the enzyme from binding to its primary DNA recognition site. Direct interaction between two protein molecules (one bound to the original recognition site and the other to a sequence-degenerated site) results in a totally new activity of PleI: it produces a nick near the degenerate site. The PNA-induced nicking efficiency varies with the distance between the two protein-binding sites in a phase with the DNA helical periodicity. Our findings imply a general approach for the fine-tuning of proteins bound to DNA sites well separated along the DNA chain. |
| Databáze: | OpenAIRE |
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