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The function of the encapsulin systems is related to the encapsulated cargo proteins. The Myxococcus xanthus encapsulin (EncA) naturally encapsulates ferritin-like proteins EncB and EncC as cargo, resulting in a large iron storage nanocompartment, able to accommodate up to 30,000 iron atoms per shell. In the present manuscript we describe the binding and protection of circular double stranded DNA (pUC19) by EncA using electrophoretic mobility shift assays (EMSA), atomic force microscopy (AFM), and DNase protection assays. EncA binds pUC19 with an apparent dissociation constant of 0.3 \u00b1 0.1 \u00b5M and a Hill coefficient of 1.4 \u00b1 0.1, while EncC alone showed no interaction with DNA. Accordingly, the EncAC complex displayed a similar DNA binding capacity as the EncA protein. The data suggest that initially, EncA converts the plasmid DNA from a supercoiled to a more relaxed form with a beads-on-a-string morphology. At higher concentrations, EncA self-aggregates, condensing the DNA. This process physically protects DNA from enzymatic digestion by DNase I. The secondary structure and thermal stability of EncA and the EncA\u2212pUC19 complex were evaluated using synchrotron radiation circular dichroism (SRCD) spectroscopy. The overall secondary structure of EncA is maintained upon interaction with pUC19 while the melting temperature of the protein (Tm) slightly increased from 76 \u00b1 1 \u00b0C to 79 \u00b1 1 \u00b0C. Our work reports, for the first time, the in vitro capacity of an encapsulin shell to interact and protect plasmid DNA similarly to other protein nanocages that may be relevant in vivo.<\/jats:p>","DOI":"10.3390\/ijms23147829","type":"journal-article","created":{"date-parts":[[2022,7,17]],"date-time":"2022-07-17T22:57:06Z","timestamp":1658098626000},"page":"7829","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin: A Novel Function"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3235-4183","authenticated-orcid":false,"given":"Ana V.","family":"Almeida","sequence":"first","affiliation":[{"name":"UCIBIO-Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"},{"name":"Associate Laboratory i4HB-Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"given":"Ana J.","family":"Carvalho","sequence":"additional","affiliation":[{"name":"UCIBIO-Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"},{"name":"Associate Laboratory i4HB-Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6798-111X","authenticated-orcid":false,"given":"Tom\u00e1s","family":"Calmeiro","sequence":"additional","affiliation":[{"name":"CENIMAT\/i3N, Department of Materials Science, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"given":"Nykola C.","family":"Jones","sequence":"additional","affiliation":[{"name":"ISA, Department of Physics and Astronomy, Aarhus University, 8000 Aarhus C, Denmark"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8018-5433","authenticated-orcid":false,"given":"S\u00f8ren V.","family":"Hoffmann","sequence":"additional","affiliation":[{"name":"ISA, Department of Physics and Astronomy, Aarhus University, 8000 Aarhus C, Denmark"}]},{"given":"Elvira","family":"Fortunato","sequence":"additional","affiliation":[{"name":"CENIMAT\/i3N, Department of Materials Science, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5567-6073","authenticated-orcid":false,"given":"Alice S.","family":"Pereira","sequence":"additional","affiliation":[{"name":"UCIBIO-Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"},{"name":"Associate Laboratory i4HB-Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7398-2661","authenticated-orcid":false,"given":"Pedro","family":"Tavares","sequence":"additional","affiliation":[{"name":"UCIBIO-Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"},{"name":"Associate Laboratory i4HB-Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1126\/science.1184318","article-title":"Some enzymes just need a space of their own","volume":"327","author":"Kang","year":"2010","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"214188","DOI":"10.1016\/j.ccr.2021.214188","article-title":"Encapsulin nanocages: Protein encapsulation and iron sequestration","volume":"448","author":"Almeida","year":"2021","journal-title":"Coord. 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