3D Stimulated Emission Depletion super resolution microscopy reveals novel association between replication factories and organized smooth tubular matrices during Zika virus infection
department or school: Medicine, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Long, Rory Kirk Mackenzie
issued: 2021-01-31T08:00:00Z
issued: 2019
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: The Zika virus (ZIKV) is a positive-sense RNA flavivirus that has been determined as a causative agent of severe neurological diseases including Guillain-Barré syndrome and microcephaly. A common characteristic of flavivirus infection is the re-organization of host cell membranes, usually endoplasmic reticulum (ER) derived, to promote viral replication. As ZIKV-induced ER structures, being vesicular replication factories (RF) and convoluted membranes (CM), are smaller than the diffraction limit of light, studies to characterize these structures and their protein compositions have been limited. In this thesis, Stimulated Emission Depletion (STED) super resolution microscopy is utilized to study the whole cell organization of ZIKV-induced ER morphologies and several viral proteins involved in replication. Generally in cells, the ER can be divided into two regions, central ER sheets and peripheral ER tubules. Here we report the formation of an ZIKV-induced, organized crescent-shaped, dense central ER region of ER tubules. ZIKV RFs localize to this dense, central ER region in a similar crescent-shaped organization. Computational 3D reconstruction of these 3D STED imaged ZIKV-induced ER structures, along with validation by electron microscopy of ZIKV infected cerebral organoids, revealed that these dense ER regions are composed of novel smooth ER tubular matrices. Moreover, we have found that ZIKV NS4B, a viral encoded integral membrane protein predicted to play a role in ER membrane re-organization during ZIKV replication, is enriched in ZIKV-induced tubular matrices. Additionally, a subpopulation of ZIKV NS4B localizes to active RFs suggesting that NS4B plays a role in the formation of both ZIKV-induced tubular matrices and RFs among other functions. Overall, the application of super resolution microscopy to study ZIKV replication has led to the identification of novel ZIKV-induced, RF-containing tubular matrices and characterization of a viral protein involved in this…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/73090
note: 3D Stimulated Emission Depletion super resolution microscopy reveals novel association between replication factories and organized smooth tubular matrices during Zika virus infection by Rory Kirk Mackenzie Long B.Sc., The University of British Columbia, 2017 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Cell and Developmental Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) December 2019 © Rory Kirk Mackenzie Long, 2019 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: 3D Stimulated Emission Depletion super resolution microscopy reveals novel association between replication factories and organized smooth tubular matrices during Zika virus infection submitted by Rory Long in partial fulfillment of the requirements for the degree of Master of Science in Cell & Developmental Biology Examining Committee: Robert Nabi, Cellular & Physiological Sciences Co-supervisor Francois Jean, Microbiology and Immunology Co-supervisor Supervisory Committee Member Lacey Samuels, Botany Additional Examiner Additional Supervisory Committee Members: Wayne Vogl, Cellular & Physiological Sciences Supervisory Committee Member Eric Jan, Biochemistry and Molecular Sciences Supervisory Committee Member iii Abstract The Zika virus (ZIKV) is a positive-sense RNA flavivirus that has been determined as a causative agent of severe neurological diseases including Guillain-Barré syndrome and microcephaly. A common characteristic of flavivirus infection is the re-organization of host cell membranes, usually endoplasmic reticulum (ER) derived, to promote viral replication. As ZIKV-induced ER structures, being vesicular replication factories (RF) and convoluted membranes (CM), are smaller than the diffraction limit of light, studies to characterize these structures and their protein co…
has type: Thesis/Dissertation
date issued: 2020-05
is shown at: 10.14288/1.0387433
language: eng
degree discipline: Cell and Developmental Biology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/73090