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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, Steve
dc.date.accessioned2020-07-16T23:30:21Z
dc.date.available2020-07-16T23:30:21Z
dc.date.issued2020
dc.identifier.issn1932-6203
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/308027
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.format.mediumElectronic-eCollection
dc.languageeng
dc.publisherPublic Library of Science (PLoS)
dc.rightsAll rights reserved
dc.subjectAnimals
dc.subjectDrosophila melanogaster
dc.subjectIsoenzymes
dc.subjectGlycogen Synthase Kinase 3
dc.subjectDrosophila Proteins
dc.subjectProteomics
dc.titleCharacterisation of protein isoforms encoded by the Drosophila Glycogen Synthase Kinase 3 gene shaggy.
dc.typeArticle
prism.issueIdentifier8
prism.publicationDate2020
prism.publicationNamePLoS One
prism.startingPagee0236679
prism.volume15
dc.identifier.doi10.17863/CAM.55122
dcterms.dateAccepted2020-07-09
rioxxterms.versionofrecord10.1371/journal.pone.0236679
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2020-01
dc.contributor.orcidKorona, Dagmara [0000-0002-5988-3894]
dc.contributor.orcidFischer, Bettina [0000-0003-2821-6287]
dc.contributor.orcidLilley, Kathryn [0000-0003-0594-6543]
dc.contributor.orcidRussell, Steve [0000-0003-0546-3031]
dc.identifier.eissn1932-6203
rioxxterms.typeJournal Article/Review
pubs.funder-project-idBBSRC (via University of Manchester) (R116574-LLM)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/L002817/1)
cam.issuedOnline2020-08-06
cam.orpheus.counter4
rioxxterms.freetoread.startdate2023-07-16


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