Characterisation of protein isoforms encoded by the Drosophila Glycogen Synthase Kinase 3 gene shaggy
Authors
Nightingale, Daniel
Fabre, Bertrand
Nelson, Michael
Fischer, Bettina
Johnson, Glynnis
Lees, Jonathan
Hubbard, Simon
Lilley, Kathryn
Publication Date
2020-08-06Journal Title
PLOS ONE
Publisher
Public Library of Science
Volume
15
Issue
8
Language
en
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Korona, D., Nightingale, D., Fabre, B., Nelson, M., Fischer, B., Johnson, G., Lees, J., et al. (2020). Characterisation of protein isoforms encoded by the Drosophila Glycogen Synthase Kinase 3 gene shaggy. PLOS ONE, 15 (8)https://doi.org/10.1371/journal.pone.0236679
Abstract
The Drosophila shaggy gene (sgg, GSK-3) encodes multiple protein isoforms with serine/threonine kinase activity and is a key player in diverse developmental signalling pathways. Currently it is unclear whether different Sgg proteoforms are similarly involved in signalling or if different proteoforms have distinct functions. We used CRISPR/Cas9 genome engineering to tag eight different Sgg proteoform classes and determined their localization during embryonic development. We performed proteomic analysis of the two major proteoform classes and generated mutant lines for both of these for transcriptomic and phenotypic analysis. We uncovered distinct tissue-specific localization patterns for all of the tagged proteoforms we examined, most of which have not previously been characterised directly at the protein level, including one proteoform initiating with a non-standard codon. Collectively, this suggests complex developmentally regulated splicing of the sgg primary transcript. Further, affinity purification followed by mass spectrometric analyses indicate a different repertoire of interacting proteins for the two major proteoforms we examined, one with ubiquitous expression (Sgg-PB) and one with nervous system specific expression (Sgg-PA). Specific mutation of these proteoforms shows that Sgg-PB performs the well characterised maternal and zygotic segmentations functions of the sgg locus, while Sgg-PA mutants show adult lifespan and locomotor defects consistent with its nervous system localisation. Our findings provide new insights into the role of GSK-3 proteoforms and intriguing links with the GSK-3α and GSK-3β proteins encoded by independent vertebrate genes. Our analysis suggests that different proteoforms generated by alternative splicing are likely to perform distinct functions.
Keywords
Research Article, Research and analysis methods, Biology and life sciences, Medicine and health sciences
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/L002817/1)
Identifiers
pone-d-20-12754
External DOI: https://doi.org/10.1371/journal.pone.0236679
This record's URL: https://www.repository.cam.ac.uk/handle/1810/308868
Rights
Licence:
http://creativecommons.org/licenses/by/4.0/