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Characterising mutant Cyclin F in the development of amyotrophic lateral sclerosis

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thesis
posted on 28.03.2022, 16:55 by Stephanie Rayner
Amyotrophic lateral sclerosis is a relentless neurodegenerative disease typically leading to death of patients within 5 years from onset. Recently, several ALS-causing gene mutations in CCNF, the gene encoding Cyclin F have been identified. We have focused on the substitution from serine to glycine at amino acid 621 and show that mutation affects the functionality of the ubiquitin ligase, SCF Cyclin F. Specifically, the mutation impacts proteostasis of a known substrate, RRM2 (ribonucleotide reductase family member 2). Although the abundance of RRM2 is not affected, there is increasing abundance of ubiquitinated RRM2 in transfected Neuro-2A cells after 24 hours post-transfection. Using a label-free proteomic workflow, a total of 60 differentially expressed proteins were associated with mutant Cyclin F-mCherry expressing cells after 8 and 24 hours post-transfection. Many changes are functionally involved in protein translation and RNA processing, which is consistent with ALS pathogenesis. Using a workflow consisting of SDS-PAGE and LC-MS/MS we also obtained protein aggregate profiles that are linked to the mutation in Neuro-2A cells under heat-stress.Notably, Septin 2 was increasingly insoluble in association with mutant Cyclin F. This is a strong candidate for validation studies, as it is known to form insoluble aggregates within Parkinsonism models.

History

Table of Contents

1. Introduction -- 2. Methods -- 3. Results -- 4. Discussion.

Notes

Bibliography: pages 54-59 Theoretical thesis.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes

Degree

MRes, Macquarie University, Faculty of Science and Engineering, Department of Chemistry and Biomolecular Sciences

Department, Centre or School

Department of Chemistry and Biomolecular Sciences

Year of Award

2015

Principal Supervisor

Mark Molloy

Additional Supervisor 1

Roger Chung

Rights

Copyright Stephanie Rayner 2014. Copyright disclaimer: http://mq.edu.au/library/copyright

Language

English

Extent

1 online resource (x. 59, 10 pages) colour illustrations

Former Identifiers

mq:53821 http://hdl.handle.net/1959.14/1137868