Date of Award
2-19-2025
Publication Type
Master Thesis
Degree Name
M.Sc.
Department
Biological Sciences
Supervisor
Lisa Porter
Rights
info:eu-repo/semantics/embargoedAccess
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Abstract
Spy1, an unconventional cyclin-like protein, serves as a potent activator of cell cycle-associated CDKs, playing crucial roles in cell cycle progression, DNA damage repair, and cancer biology. This study examines Spy1's interactions with critical regulatory proteins, including p27 and SUN1 splicing variants (SUN1_916 and SUN1_785), to better understand its functional roles. Using HEK293 cells transfected with wild-type and mutant Spy1 constructs, we investigated the impact of conserved residues within Spy1’s S/R box on its ability to bind p27. While prior studies reported a significant effect of Spy1 mutations on p27 turnover, our results showed no notable changes in p27 levels. The binding dynamics between Spy1 and p27 were further assessed through coimmunoprecipitation assays, revealing no significant differences in p27 binding among Spy1 mutants. These discrepancies with earlier findings underscore the importance of consistent methodologies to clarify Spy1-p27 interactions. Moreover, analysis of the CDK2 inhibitor NU6102 showed no significant impact on p27 level in the presence of Spy1, highlighting the complexity of cell cycle regulation. We also investigated Spy1's interaction with SUN1 splicing variants, finding that Spy1 displayed a stronger affinity for SUN1_916 compared to the shorter SUN1_785 isoform. Further evaluation displayed that Both SUN1 variants colocalized with Spy1 at the nuclear envelope, but their overexpression did not significantly affect cell proliferation or cell cycle profiles. These findings underscore Spy1's multifaceted roles and potential as a therapeutic target, highlighting the need for further research into its downstream pathways and interactions.
Recommended Citation
Salimi Chirani, Alireza, "Elucidating the Interactions of Spy1 with p27 and SUN1 Splice Variants in Cell Cycle Regulation" (2025). Electronic Theses and Dissertations. 9687.
https://scholar.uwindsor.ca/etd/9687
