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Viral Capsid Change upon Encapsulation of Double-Stranded DNA into an Infectious Hypodermal and Hematopoietic Necrosis Virus-like Particle.
Weerachatyanukul, Wattana; Kiatmetha, Pauline; Raksat, Ponlawoot; Boonkua, Supawich; Thongsum, Orawan; Jariyapong, Pitchanee; Chotwiwatthanakun, Charoonroj; Ounjai, Puey; Metlagel, Zoltan.
Afiliación
  • Weerachatyanukul W; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Ratchathewi, Bangkok 10400, Thailand.
  • Kiatmetha P; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Ratchathewi, Bangkok 10400, Thailand.
  • Raksat P; Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok 10400, Thailand.
  • Boonkua S; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Ratchathewi, Bangkok 10400, Thailand.
  • Thongsum O; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Ratchathewi, Bangkok 10400, Thailand.
  • Jariyapong P; Department of Medical Science, School of Medicine, Walailak University, Thasala District, Nakhonsrithammarat 80160, Thailand.
  • Chotwiwatthanakun C; Academic and Curriculum Division, Nakhonsawan Campus, Mahidol University, Nakhonsawan 60130, Thailand.
  • Ounjai P; Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok 10400, Thailand.
  • Metlagel Z; Lawrence Berkeley National Laboratory, Molecular Biophysics and Integrated Bioimaging Division, University of California, Berkeley, CA 94720, USA.
Viruses ; 15(1)2022 12 30.
Article en En | MEDLINE | ID: mdl-36680151
In this study, we aimed to encapsulate the sizable double-stranded DNA (dsDNA, 3.9 kbp) into a small-sized infectious hypodermal and hematopoietic necrosis virus-like particle (IHHNV-VLP; T = 1) and compared the changes in capsid structure between dsDNA-filled VLP and empty VLP. Based on our encapsulation protocol, IHHNV-VLP was able to load dsDNA at an efficiency of 30-40% (w/w) into its cavity. Structural analysis revealed two subclasses of IHHNV-VLP, so-called empty and dsDNA-filled VLPs. The three-dimensional (3D) structure of the empty VLP produced in E. coli was similar to that of the empty IHHNV-VLP produced in Sf9 insect cells. The size of the dsDNA-filled VLP was slightly bigger (50 Å) than its empty VLP counterpart; however, the capsid structure was drastically altered. The capsid was about 1.5-fold thicker due to the thickening of the capsid interior, presumably from DNA-capsid interaction evident from capsid protrusions or nodules on the interior surface. In addition, the morphological changes of the capsid exterior were particularly observed in the vicinity of the five-fold axes, where the counter-clockwise twisting of the "tripod" structure at the vertex of the five-fold channel was evident, resulting in a widening of the channel's opening. Whether these capsid changes are similar to virion capsid maturation in the host cells remains to be investigated. Nevertheless, the ability of IHHNV-VLP to encapsulate the sizable dsDNA has opened up the opportunity to package a dsDNA vector that can insert exogenous genes and target susceptible shrimp cells in order to halt viral infection.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cápside / Densovirinae Idioma: En Revista: Viruses Año: 2022 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Suiza
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cápside / Densovirinae Idioma: En Revista: Viruses Año: 2022 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Suiza