Evaluation of the Role of Bacterial Amyloid on Nucleoid Structure Using Cryo-Soft X-Ray Tomography.

Autor:	Cossa A; Laboratoire Léon Brillouin LLB, CEA, CNRS UMR 12, Université Paris-Saclay, CEA Saclay, Gif-sur-Yvette, France.; Institut Curie, Université PSL, CNRS UAR2016, Inserm US43, Université Paris-Saclay, Multimodal Imaging Center, Orsay, France.; National Center of Biotechnology, CSIC, Campus Univ. Autónoma de Madrid, Madrid, Spain., Wien F; Synchrotron SOLEIL, L'Orme des Merisiers Saint Aubin, Gif-sur-Yvette, France., Turbant F; Laboratoire Léon Brillouin LLB, CEA, CNRS UMR 12, Université Paris-Saclay, CEA Saclay, Gif-sur-Yvette, France.; Department of Molecular Biology, Faculty of Biology, University of Gdansk, Gdansk, Poland., Kaczorowski T; Department of Microbiology, Faculty of Biology, University of Gdansk, Gdansk, Poland., Węgrzyn G; Department of Molecular Biology, Faculty of Biology, University of Gdansk, Gdansk, Poland., Arluison V; Laboratoire Léon Brillouin LLB, CEA, CNRS UMR 12, Université Paris-Saclay, CEA Saclay, Gif-sur-Yvette, France.; Université de Paris Cité, Paris, France., Pérez-Berná AJ; Mistral Beamline, Alba Light Source, Barcelona, Spain., Trépout S; Institut Curie, Université PSL, CNRS UAR2016, Inserm US43, Université Paris-Saclay, Multimodal Imaging Center, Orsay, France., Pereiro E; Mistral Beamline, Alba Light Source, Barcelona, Spain. epereiro@cells.es.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2538, pp. 319-333.
DOI:	10.1007/978-1-0716-2529-3_21
Abstrakt:	Bacterial chromosomal DNA is packed within a non-membranous structure, the nucleoid, thanks to nucleoid associated proteins (NAPs). The role of bacterial amyloid has recently emerged among these NAPs, particularly with the nucleoid-associated protein Hfq that plays a direct role in DNA compaction. In this chapter, we present a 3D imaging technique, cryo-soft X-ray tomography (cryo-SXT) to obtain a detailed 3D visualization of subcellular bacterial structures, especially the nucleoid. Cryo-SXT imaging of native unlabeled cells enables observation of the nucleoid in 3D with a high resolution, allowing to evidence in vivo the role of amyloids on DNA compaction. The precise experimental methods to obtain 3D tomograms will be presented. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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