Assessment of potential :1 programmed ribosomal frameshifting by the human cytokine receptor IL-7Rα
department or school: Science, Faculty of
department or school: Microbiology and Immunology, Department of
data provider: DSpace
degree campus: UBCV
creator: Jackson, Jennie Lynne
issued: 2025-04-25T14:53:54Z
issued: 2025
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Translation is an important step in the flow of genetic information from DNA, to RNA, to protein. Yet, translational fidelity is not always maintained. One example of a recoding event is programmed ribosomal frameshifts (PRF). PRF is mainly utilized by viruses, yeast, or bacteria, where the outcome of a shifted reading frame may result in carboxy-terminally extended fusion proteins or mRNA degradation. In this way, PRF is a form of translational control. The regulation of cytokine receptors, including at the translational level, is valuable to understand due to their importance in immune cell signaling. Interleukin-7 receptor α (IL-7Rα) in particular, which has major roles in proper immune system development and function, is known to be tightly regulated. IL-7Rα has well-documented examples of transcriptional and post-translational regulation, but post-transcriptional and translational regulation remain areas requiring further study. Indeed, IL-7Rα, as well as IL-2Rγ – the other subunit of the heterodimeric IL-7 receptor – were computationally predicted to contain -1 PRF motifs in their mRNA. Initial in vitro testing with reporter constructs expressed in HeLa cells was promising, but HeLa cells are not the best model for intrinsic IL-7Rα signaling pathways. Therefore, I sought to test the putative IL-7Rα PRF motif in a physiologically relevant context. I hypothesized that translational regulation of the cytokine receptor IL-7Rα via -1 PRF modulated its expression in human T cells. Using flow cytometry, I developed a single cell level assay so that -1 PRF could be compared between different T cell subsets, such as naïve, effector, or memory, within the same sample. Through various techniques, I also characterized impaired and/or abnormal Renilla luciferase expression by our IL-7Rα PRF dual luciferase reporter plasmid. Overall, despite being unable to resolve whether IL-7Rα expression is regulated by -1 PRF in human T cells, the flow cytometry PRF assay remains an ada…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/90793
note: ASSESSMENT OF POTENTIAL -1 PROGRAMMED RIBOSOMAL FRAMESHIFTING BY THE HUMAN CYTOKINE RECEPTOR IL-7Rα by Jennie Lynne Jackson B.Sc., The University of British Columbia, 2019 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Microbiology and Immunology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) April 2025 © Jennie Lynne Jackson, 2025 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: Assessment of potential -1 programmed ribosomal frameshifting by the human cytokine receptor IL-7Rα submitted by Jennie Lynne Jackson in partial fulfilment of the requirements for the degree of Master of Science in Microbiology and Immunology Examining Committee: Dr. Ninan Abraham, Professor, Department of Microbiology and Immunology, Department of Zoology, UBC Supervisor Dr. Eric Jan, Professor, Department of Biochemistry and Molecular Biology, UBC Supervisory Committee Member Dr. Maria Tokuyama, Assistant Professor, Department of Microbiology and Immunology, UBC Additional Examiner Additional Supervisory Committee Members: Dr. François Jean, Professor, Department of Microbiology and Immunology, UBC Supervisory Committee Member iii Abstract Translation is an important step in the flow of genetic information from DNA, to RNA, to protein. Yet, translational fidelity is not always maintained. One example of a recoding event is programmed ribosomal frameshifts (PRF). PRF is mainly utilized by viruses, yeast, or bacteria, where the outcome of a shifted reading frame may result in carboxy-terminally extended fusion proteins or mRNA degradation. In this way, PRF is a form of translational control. The regulation of cytokine receptors, including at the translational level, is valuable to understand due to their importance in immune cell signaling. Interleukin-7 receptor α (IL-7Rα) in …
has type: Thesis/Dissertation
date issued: 2025-05
is shown at: 10.14288/1.0448588
language: eng
degree discipline: Microbiology and Immunology
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
contributor: Abraham, Ninan
type: Text
identifier URI: http://hdl.handle.net/2429/90793