Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2023
Höfundar / Authors: Isabelle Kristín Tarbox, Elisabet Guðmundsdóttir, Zophonías O. Jónsson and Sigríður Rut Franzdóttir
Starfsvettvangur / Affiliations: 1. Háskóli Íslands
Kynnir / Presenter: Elísabet Guðmundsdóttir and Isabelle Kristín Tarbox
The proteasome is a highly conserved complex responsible for degradation of misfolded and ubiquitinated proteins in eukaryotic cells. Failure of proteasome function is associated with many neurological diseases such as Alzheimer’s disease.
We are interested in studying how two highly conserved candidate proteins affect the performance of the proteasome, i.e., maintenance of proteasome function and response to failure in the Drosophila melanogaster nervous system. To study this, a simple fluorescent reporter system for monitoring proteasome efficiency in vivo was set up and tested, followed by experiments testing the effect of loss of the candidates on proteasome activity. A second system based on a read-out for heat-induced proteasome failure was optimized and experiments performed to test the ability of our candidates to modify the phenotype in photoreceptor neurons. To be able to visualize proteasomes in vivo, donor and gRNA constructs for endogenous tagging of three proteasome subunits with fluorescent proteins were constructed and gRNA efficiency tested in microinjected embryos via T7 endonuclease assay. Results indicate viable use of the tested gRNAs, and the reporter system provided an effective method for monitoring proteasome function. Furthermore, over-expression of the candidates yields a subtle rescue effect upon proteasome failure. Together these projects yielded valuable tools and baseline data for further study of the candidate genes and proteasomes in vivo.