IS911 transpososome assembly as analysed by tethered particle motion
| Autor: | Laurence Salomé, Michael Chandler, N. Pouget, Catherine Turlan, Nicolas Destainville |
|---|---|
| Přispěvatelé: | Laboratoire de microbiologie et génétique moléculaires (LMGM), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Physique Statistique des Systèmes Complexes (LPT) (PhyStat), Laboratoire de Physique Théorique (LPT), Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Institut de pharmacologie et de biologie structurale (IPBS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse) |
| Jazyk: | angličtina |
| Rok vydání: | 2006 |
| Předmět: |
Transposable element
DNA Bacterial MESH: Motion MESH: Microspheres Transposases MESH: Microscopy Fluorescence Biology 03 medical and health sciences chemistry.chemical_compound Motion 0302 clinical medicine Structural Biology Genetics A-DNA Electrophoretic mobility shift assay [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Microscopy Interference Binding site MESH: Microscopy Interference Transposase ComputingMilieux_MISCELLANEOUS 030304 developmental biology Repetitive Sequences Nucleic Acid [PHYS]Physics [physics] 0303 health sciences MESH: Repetitive Sequences Nucleic Acid Binding Sites Microscopy Video MESH: Kinetics Synapsis MESH: DNA Bacterial Microspheres MESH: Microscopy Video Kinetics MESH: Nucleic Acid Conformation Nucleoproteins Biochemistry Tethered particle motion chemistry MESH: Binding Sites MESH: DNA Transposable Elements Microscopy Fluorescence Biophysics MESH: Nucleoproteins DNA Transposable Elements MESH: Transposases Nucleic Acid Conformation 030217 neurology & neurosurgery DNA |
| Zdroj: | Nucleic Acids Research Nucleic Acids Research, Oxford University Press, 2006, 34, pp.4313-4323. ⟨10.1093/nar/gkl420⟩ Nucleic Acids Research, Oxford University Press, 2006, 34 (16), pp.4313-23. ⟨10.1093/nar/gkl420⟩ |
| ISSN: | 0305-1048 1362-4962 |
| DOI: | 10.1093/nar/gkl420⟩ |
| Popis: | International audience; Initiation of transposition requires formation of a synaptic complex between both transposon ends and the transposase (Tpase), the enzyme which catalyses DNA cleavage and strand transfer and which ensures transposon mobility. We have used a single-molecule approach, tethered particle motion (TPM), to observe binding of a Tpase derivative, OrfAB[149], amputated for its C-terminal catalytic domain, to DNA molecules carrying one or two IS911 ends. Binding of OrfAB[149] to a single IS911 end provoked a small shortening of the DNA. This is consistent with a DNA bend introduced by protein binding to a single end. This was confirmed using a classic gel retardation assay with circularly permuted DNA substrates. When two ends were present on the tethered DNA in their natural, inverted, configuration, Tpase not only provoked the short reduction in length but also generated species with greatly reduce effective length consistent with DNA looping between the ends. Once formed, this 'looped' species was very stable. Kinetic analysis in real-time suggested that passage from the bound unlooped to the looped state could involve another species of intermediate length in which both transposon ends are bound. DNA carrying directly repeated ends also gave rise to the looped species but the level of the intermediate species was significantly enhanced. Its accumulation could reflect a less favourable synapse formation from this configuration than for the inverted ends. This is compatible with a model in which Tpase binds separately to and bends each end (the intermediate species) and protein-protein interactions then lead to synapsis (the looped species). |
| Databáze: | OpenAIRE |
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