2026-06-14T00:27:22Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/144042024-10-31T11:53:54Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609
Repisalud
author
Patel, Nikesh
author
Clark, Sam
author
Weiß, Eva U
author
Pérez Mata, Carlos
author
Bohon, Jen
author
Farquhar, Erik R
author
Maskell, Daniel P
author
Ranson, Neil A
author
Twarock, Reidun
author
Stockley, Peter G
funder
UK Research and Innovation
funder
Medical Research Council (Reino Unido)
funder
Wellcome Trust
funder
Engineering and Physical Sciences Research Council (Reino Unido)
funder
Royal Society (Reino Unido)
funder
National Science Foundation (Estados Unidos)
funder
National Institutes of Health (Estados Unidos)
funder
United States Department of Energy
funder
University of Leeds (Reino Unido)
funder
Unión Europea. Comisión Europea
2022-05-17T11:11:18Z
2022-05-17T11:11:18Z
2021-12-16
Commun Biol. 2021 Dec 16;4(1):1407.
10.1038/s42003-021-02897-2
2399-3642
Communications Biology
34916604
http://hdl.handle.net/20.500.12105/14404
The roles of RNA sequence/structure motifs, Packaging Signals (PSs), for regulating assembly of an HBV genome transcript have been investigated in an efficient in vitro assay containing only core protein (Cp) and RNA. Variants of three conserved PSs, within the genome of a strain not used previously, preventing correct presentation of a Cp-recognition loop motif are differentially deleterious for assembly of nucleocapsid-like particles (NCPs). Cryo-electron microscopy reconstruction of the T = 4 NCPs formed with the wild-type gRNA transcript, reveal that the interior of the Cp shell is in contact with lower resolution density, potentially encompassing the arginine-rich protein domains and gRNA. Symmetry relaxation followed by asymmetric reconstruction reveal that such contacts are made at every symmetry axis. We infer from their regulation of assembly that some of these contacts would involve gRNA PSs, and confirmed this by X-ray RNA footprinting. Mutation of the ε stem-loop in the gRNA, where polymerase binds in vivo, produces a poor RNA assembly substrate with Cp alone, largely due to alterations in its conformation. The results show that RNA PSs regulate assembly of HBV genomic transcripts in vitro, and therefore may play similar roles in vivo, in concert with other molecular factors.
eng
In vitro functional analysis of gRNA sites regulating assembly of hepatitis B virus
journal article
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