Real-time compaction of nanoconfined DNA by an intrinsically disordered macromolecular counterion.
Biochem Biophys Res Commun
; 533(1): 175-180, 2020 11 26.
Article
em En
| MEDLINE
| ID: mdl-32951838
ABSTRACT
We demonstrate how a recently developed nanofluidic device can be used to study protein-induced compaction of genome-length DNA freely suspended in solution. The protein we use in this study is the hepatitis C virus core protein (HCVcp), which is a positively charged, intrinsically disordered protein. Using nanofluidic devices in combination with fluorescence microscopy, we observe that protein-induced compaction preferentially begins at the ends of linear DNA. This observation would be difficult to make with many other single-molecule techniques, which generally require the DNA ends to be anchored to a substrate. We also demonstrate that this protein-induced compaction is reversible and can be dynamically modulated by exposing the confined DNA molecules to solutions containing either HCVcp (to promote compaction) or Proteinase K (to disassemble the compact nucleo-protein complex). Although the natural binding partner for HCVcp is genomic viral RNA, the general biophysical principles governing protein-induced compaction of DNA are likely relevant for a broad range of nucleic acid-binding proteins and their targets.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
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Proteínas do Core Viral
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Hepacivirus
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Dispositivos Lab-On-A-Chip
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Proteínas Intrinsicamente Desordenadas
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article