eprintid: 10053748
rev_number: 26
eprint_status: archive
userid: 608
dir: disk0/10/05/37/48
datestamp: 2018-08-08 12:01:45
lastmod: 2021-08-02 22:42:28
status_changed: 2018-08-08 12:01:45
type: thesis
metadata_visibility: show
creators_name: Donaldson, Callum
title: Investigating the relationship between core stability and early life cycle events in HIV-1
ispublished: unpub
divisions: UCL
divisions: A01
divisions: B02
divisions: C10
abstract: HIV-1 capsid (CA) plays a vital role in the early stages of HIV-1 infection. The CA lattice surrounding the viral core is predominantly assembled from CA hexamers and stabilised by intra-hexamer and inter-hexamer interactions. Optimal stability of the lattice is known to be critical for efficient infection; however, a comprehensive screen of the effects of stabilising all lattice interfaces has not been performed. Disulphide cross-linking of residues across lattice interfaces has been used in vitro to stabilise CA assemblies. In this study, putatively stabilising cysteine CA mutations were designed at each interface of the CA lattice and their effects on early life cycle events, including reverse transcription and nuclear entry, assessed. The introduction of cysteine mutations at intra-hexamer (both NTD-NTD and NTD-CTD) and inter-hexamer (dimeric CTD-CTD only) lattice interfaces resulted in cross-linking and hyperstable viral cores in infected cells. These cores were minimally infectious and encountered sequential blocks to infectivity at reverse transcription, nuclear entry and post-nuclear entry. The infectivity defect of hyper-stable core mutant, A14C/E45C, was partially compensated – without an observable decrease in stability – by addition of mutations reported to perturb interactions with CPSF6. In contrast, Nup153 and CypA mutations were unable to compensate the infectivity defect suggesting that this was a CPSF6-specific effect. Proximal ligation assays were performed to visualise and quantify interactions between CA and host factors, indicating that hyper-stable cores encountered a block to nuclear entry in G1/S arrested cells. Overall, the results of this study suggest that mutations at different lattice interfaces can result in global changes to the intrinsic stability of the viral core and results in fitness defects at multiple stages of the HIV-1 early life cycle.
date: 2018-07-28
date_type: published
oa_status: green
full_text_type: other
thesis_class: doctoral_open
thesis_award: Ph.D
language: eng
thesis_view: UCL_Thesis
primo: open
primo_central: open_green
verified: verified_manual
elements_id: 1573223
lyricists_name: Donaldson, Callum
lyricists_id: CDDON02
actors_name: Donaldson, Callum
actors_name: Allington-Smith, Dominic
actors_id: CDDON02
actors_id: DAALL44
actors_role: owner
actors_role: impersonator
full_text_status: public
pages: 227
event_title: UCL (University College London)
institution: UCL (University College London)
department: Department of Infection and Immunity
thesis_type: Doctoral
citation:        Donaldson, Callum;      (2018)    Investigating the relationship between core stability and early life cycle events in HIV-1.                   Doctoral thesis  (Ph.D), UCL (University College London).     Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10053748/1/Donaldson_10053748_thesis.pdf