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dc.contributor.authorKorona, Dagmara
dc.contributor.authorNightingale, Daniel
dc.contributor.authorFabre, Bertrand
dc.contributor.authorNelson, Michael
dc.contributor.authorFischer, Bettina
dc.contributor.authorJohnson, Glynnis
dc.contributor.authorLees, Jonathan
dc.contributor.authorHubbard, Simon
dc.contributor.authorLilley, Kathryn
dc.contributor.authorRussell, Steven
dc.date.accessioned2020-08-07T01:09:32Z
dc.date.available2020-08-07T01:09:32Z
dc.date.issued2020-08-06
dc.date.submitted2020-05-01
dc.identifier.otherpone-d-20-12754
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/308868
dc.description.abstractThe 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.
dc.languageen
dc.publisherPublic Library of Science
dc.subjectResearch Article
dc.subjectResearch and analysis methods
dc.subjectBiology and life sciences
dc.subjectMedicine and health sciences
dc.titleCharacterisation of protein isoforms encoded by the Drosophila Glycogen Synthase Kinase 3 gene shaggy
dc.typeArticle
dc.date.updated2020-08-07T01:09:31Z
prism.issueIdentifier8
prism.publicationNamePLOS ONE
prism.volume15
dc.identifier.doi10.17863/CAM.55958
dcterms.dateAccepted2020-07-09
rioxxterms.versionofrecord10.1371/journal.pone.0236679
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
datacite.contributor.supervisoreditor: Klymkowsky, Michael
dc.contributor.orcidKorona, Dagmara [0000-0002-5988-3894]
dc.contributor.orcidRussell, Steven [0000-0003-0546-3031]
dc.identifier.eissn1932-6203
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/L002817/1)


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