Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin

Detalhes bibliográficos
Autor(a) principal: Almeida, Ana V.
Data de Publicação: 2022
Outros Autores: Carvalho, Ana J., Calmeiro, Tomás, Jones, Nykola C., Hoffmann, Søren V., Fortunato, Elvira, Pereira, Alice S., Tavares, Pedro
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/150712
Resumo: Funding Information: This work was supported by Fundação para a Ciência e Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES), through the Radiation Biology and Biophysics Doctoral Training Programme—RaBBiT (PD/00193/2012). This work benefited from STSM funding by COST Action (CA15126 MOBIEU). Publisher Copyright: © 2022 by the authors.
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spelling Condensation and Protection of DNA by the Myxococcus xanthus EncapsulinA Novel FunctionAFMDNA binding and protectionDNA condensationEMSAencapsulinprotein nanocagesSRCDthermal stabilityCatalysisMolecular BiologySpectroscopyComputer Science ApplicationsPhysical and Theoretical ChemistryOrganic ChemistryInorganic ChemistryFunding Information: This work was supported by Fundação para a Ciência e Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES), through the Radiation Biology and Biophysics Doctoral Training Programme—RaBBiT (PD/00193/2012). This work benefited from STSM funding by COST Action (CA15126 MOBIEU). Publisher Copyright: © 2022 by the authors.Encapsulins are protein nanocages capable of harboring smaller proteins (cargo proteins) within their cavity. 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 ± 0.1 µM and a Hill coefficient of 1.4 ± 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−pUC19 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 ± 1 °C to 79 ± 1 °C. 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.DQ - Departamento de QuímicaUCIBIO - Applied Molecular Biosciences UnitCENIMAT-i3N - Centro de Investigação de Materiais (Lab. Associado I3N)DCM - Departamento de Ciência dos MateriaisRUNAlmeida, Ana V.Carvalho, Ana J.Calmeiro, TomásJones, Nykola C.Hoffmann, Søren V.Fortunato, ElviraPereira, Alice S.Tavares, Pedro2023-03-16T22:36:41Z2022-072022-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article15application/pdfhttp://hdl.handle.net/10362/150712eng1661-6596PURE: 56066372https://doi.org/10.3390/ijms23147829info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:33:01Zoai:run.unl.pt:10362/150712Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:54:16.247390Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
A Novel Function
title Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
spellingShingle Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
Almeida, Ana V.
AFM
DNA binding and protection
DNA condensation
EMSA
encapsulin
protein nanocages
SRCD
thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
title_short Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
title_full Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
title_fullStr Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
title_full_unstemmed Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
title_sort Condensation and Protection of DNA by the Myxococcus xanthus Encapsulin
author Almeida, Ana V.
author_facet Almeida, Ana V.
Carvalho, Ana J.
Calmeiro, Tomás
Jones, Nykola C.
Hoffmann, Søren V.
Fortunato, Elvira
Pereira, Alice S.
Tavares, Pedro
author_role author
author2 Carvalho, Ana J.
Calmeiro, Tomás
Jones, Nykola C.
Hoffmann, Søren V.
Fortunato, Elvira
Pereira, Alice S.
Tavares, Pedro
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv DQ - Departamento de Química
UCIBIO - Applied Molecular Biosciences Unit
CENIMAT-i3N - Centro de Investigação de Materiais (Lab. Associado I3N)
DCM - Departamento de Ciência dos Materiais
RUN
dc.contributor.author.fl_str_mv Almeida, Ana V.
Carvalho, Ana J.
Calmeiro, Tomás
Jones, Nykola C.
Hoffmann, Søren V.
Fortunato, Elvira
Pereira, Alice S.
Tavares, Pedro
dc.subject.por.fl_str_mv AFM
DNA binding and protection
DNA condensation
EMSA
encapsulin
protein nanocages
SRCD
thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
topic AFM
DNA binding and protection
DNA condensation
EMSA
encapsulin
protein nanocages
SRCD
thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
description Funding Information: This work was supported by Fundação para a Ciência e Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES), through the Radiation Biology and Biophysics Doctoral Training Programme—RaBBiT (PD/00193/2012). This work benefited from STSM funding by COST Action (CA15126 MOBIEU). Publisher Copyright: © 2022 by the authors.
publishDate 2022
dc.date.none.fl_str_mv 2022-07
2022-07-01T00:00:00Z
2023-03-16T22:36:41Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/150712
url http://hdl.handle.net/10362/150712
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1661-6596
PURE: 56066372
https://doi.org/10.3390/ijms23147829
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 15
application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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