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A novel role for gag as a cis-acting element regulating RNA structure, dimerization and packaging in HIV-1 lentiviral vectors.

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cam.issuedOnline2021-12-20
cam.orpheus.successTue Feb 01 19:02:20 GMT 2022 - Embargo updated*
dc.contributor.authorVamva, Eirini
dc.contributor.authorGriffiths, Alex
dc.contributor.authorVink, Conrad A
dc.contributor.authorLever, Andrew
dc.contributor.authorKenyon, Julia
dc.contributor.orcidVink, Conrad A [0000-0001-6739-6669]
dc.contributor.orcidKenyon, Julia [0000-0001-6055-7052]
dc.date.accessioned2021-11-25T00:31:14Z
dc.date.available2021-11-25T00:31:14Z
dc.date.issued2022-01-11
dc.description.abstractClinical usage of lentiviral vectors is now established and increasing but remains constrained by vector titer with RNA packaging being a limiting factor. Lentiviral vector RNA is packaged through specific recognition of the packaging signal on the RNA by the viral structural protein Gag. We investigated structurally informed modifications of the 5' leader and gag RNA sequences in which the extended packaging signal lies, to attempt to enhance the packaging process by facilitating vector RNA dimerization, a process closely linked to packaging. We used in-gel SHAPE to study the structures of these mutants in an attempt to derive structure-function correlations that could inform optimized vector RNA design. In-gel SHAPE of both dimeric and monomeric species of RNA revealed a previously unreported direct interaction between the U5 region of the HIV-1 leader and the downstream gag sequences. Our data suggest a structural equilibrium exists in the dimeric viral RNA between a metastable structure that includes a U5-gag interaction and a more stable structure with a U5-AUG duplex. Our data provide clarification for the previously unexplained requirement for the 5' region of gag in enhancing genomic RNA packaging and provide a basis for design of optimized HIV-1 based vectors.
dc.description.sponsorshipEV was supported by a grant from the BBSRC (BB/N503708/1 to AMLL and CV) and received personal support from Darwin College, the University of Cambridge trust and the Philosophical society of Cambridge. JCK received personal support from Homerton College. AMLL is supported by the Clinical Academic Reserve and his laboratory by the NIHR Cambridge BRC (Grant RCAG/18).
dc.identifier.doi10.17863/CAM.78519
dc.identifier.eissn1362-4962
dc.identifier.issn0305-1048
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/331074
dc.language.isoeng
dc.publisherOxford University Press (OUP)
dc.rightsAll rights reserved
dc.rights.urihttp://www.rioxx.net/licenses/all-rights-reserved
dc.titleA novel role for gag as a cis-acting element regulating RNA structure, dimerization and packaging in HIV-1 lentiviral vectors.
dc.typeArticle
dcterms.dateAccepted2021-11-30
prism.publicationNameNucleic Acids Res
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/N503708/1)
rioxxterms.licenseref.startdate2021-11-22
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionAM
rioxxterms.versionofrecord10.1093/nar/gkab1206

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