Investigation of cryptic transcription suppressing factors using a FLO8:sfGFP reporter
department or school: Medicine, Faculty of
department or school: Biochemistry and Molecular Biology, Department of
data provider: DSpace
degree campus: UBCV
creator: Gao, Xiaochen
issued: 2023-04-18T22:08:15Z
issued: 2023
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Chromatin disruption caused by polymerase passage during transcription or DNA replication has the potential to lead to cryptic transcription originating within aberrant initiation sites across the genome. We developed a sfGFP fluorescence reporter to track cryptic transcription initiating from an intragenic cryptic promoter within the FLO8 coding region and confirmed reporter utility in a spt16-197 mutant strain. Using this reporter, our results showed that the proteins, Spt10, Spt21, Cac1, and Asf1 are required for chromatin stability during DNA replication, while Spt2 plays a replication-independent role in maintaining chromatin. We also showed that blocking DNA replication only partially suppresses cryptic transcription in Spt5, Spt6, or Spt16-depleted cells, indicative of these factors functioning in a process independent of replication. Taken together, our results demonstrate that restoration of nucleosome stability following disruption is facilitated by a number of collaborating factors functioning in various pathways.
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/84313
note: INVESTIGATION OF CRYPTIC TRANSCRIPTION SUPPRESSING FACTORS USING A FLO8:sfGFP REPORTER by Xiaochen Gao 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 (Biochemistry and Molecular Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) March 2023 © Xiaochen Gao, 2023 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: Investigation of cryptic transcription suppressing factors using a FLO8:sfGFP reporter submitted by Xiaochen Gao in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry and Molecular Biology Examining Committee: LeAnn Howe, Professor, Biochemistry and Molecular Biology, UBC Supervisor Sheila Teves, Assistant Professor, Biochemistry and Molecular Biology, UBC Supervisory Committee Member Annie Ciernia, Assistant Professor, Biochemistry and Molecular Biology, UBC Supervisory Committee Member Vivien Measday, Associate Professor, Faculty of Land and Food Systems, UBC Additional Examiner iii Abstract Chromatin disruption caused by polymerase passage during transcription or DNA replication has the potential to lead to cryptic transcription originating within aberrant initiation sites across the genome. We developed a sfGFP fluorescence reporter to track cryptic transcription initiating from an intragenic cryptic promoter within the FLO8 coding region and confirmed reporter utility in a spt16-197 mutant strain. Using this reporter, our results showed that the proteins, Spt10, Spt21, Cac1, and Asf1 are required for chromatin stability during DNA replication, while Spt2 plays a replication-independent role in maintaining chromatin. We also showed that blocking DNA replication only partially suppresses cryptic transcription in Spt5, Spt6, or Spt16-depleted cells, indicati…
has type: Thesis/Dissertation
date issued: 2023-05
is shown at: 10.14288/1.0431108
language: eng
degree discipline: Biochemistry and Molecular 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
contributor: Howe, LeAnn
type: Text
identifier URI: http://hdl.handle.net/2429/84313