Nanoscale surface structures of DNA bound to Deinococcus radiodurans HU unveiled by atomic force microscopy. - Archive ouverte HAL Access content directly
Journal Articles Nanoscale Year : 2020

Nanoscale surface structures of DNA bound to Deinococcus radiodurans HU unveiled by atomic force microscopy.

Abstract

The Deinococcus radiodurans protein HU (DrHU) was shown to be critical for nucleoid activities, yet its functional and structural properties remain largely unexplored. We have applied atomic force microscopy (AFM) imaging to study DrHU binding to pUC19-DNA in vitro and analyzed the topographic structures formed at the nanoscale. At the single-molecule level, AFM imaging allows visualization of super-helical turns on naked DNA surfaces and characterization of free DrHU molecules observed as homodimers. When enhancing the molecular surface structures of AFM images by the Laplacian weight filter, the distribution of bound DrHUs was visibly varied as a function of the DrHU/DNA molar ratio. At a low molar ratio, DrHU binding was found to reduce the volume of condensed DNA configuration by about 50%. We also show that DrHU is capable of bridging distinct DNA segments. Moreover, at a low molar ratio, the binding orientation of individual DrHU dimers could be perceived on partially "open" DNA configuration. At a high molar ratio, DrHU stiffened the DNA molecule and enlarged the spread of the open DNA configuration. Furthermore, a lattice-like pattern could be seen on the surface of DrHU-DNA complex, indicating that DrHU multimerization had occurred leading to the formation of a higher order architecture. Together, our results show that the functional plasticity of DrHU in mediating DNA organization is subject to both the conformational dynamics of DNA molecules and protein abundance.
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Dates and versions

hal-03018678 , version 1 (23-11-2020)

Identifiers

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Shu-Wen W. Chen, Anne-Sophie Banneville, Jean-Marie Teulon, Joanna Timmins, Jean-Luc Pellequer. Nanoscale surface structures of DNA bound to Deinococcus radiodurans HU unveiled by atomic force microscopy.. Nanoscale, 2020, 12 (44), pp.22628-22638. ⟨10.1039/d0nr05320a⟩. ⟨hal-03018678⟩
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