Disordered Expression of shaggy, the Drosophila Gene Encoding a Serine-Threonine Protein Kinase GSK3, Affects the Lifespan in a Transcript-, Stage-, and Tissue-Specific Manner

Trostnikov, Mikhail V.; Roshina, Natalia V.; Boldyrev, S. V.; Veselkina, Ekaterina R.; Zhuikov, Andrey A; Krementsova, A. V.; Pasyukova, Elena G.

doi:10.3390/ijms20092200

Cited by 12 publications

(35 citation statements)

References 60 publications

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“…It was shown that the ubiquitous overexpression of sgg decreased lifespan and that ubiquitous sgg RNAi knockdown increased lifespan (Castillo-Quan et al, 2016). Later, we demonstrated that the misexpression of sgg affected lifespan in a transcript-, stage-, tissue-and sex-specific mode (Trostnikov et al, 2019). For example, the overexpression of the sgg-RB transcript (GenBank 2 # AY122193.1; encodes the major GSK3 isoform (Ruel et al, 1993;Bourouis, 2002), in the nervous system caused pathological changes in neurons paralleled by a rapid decline in survival and severe shortening of lifespan, whereas sgg-RB overexpression in muscles caused only a weak decrease in survival.…”

Section: Introductionmentioning

confidence: 75%

“…Lifespan was measured as described by Roshina et al (2014) and Trostnikov et al (2019). Five virgin flies of the same genotype and sex, all collected on the same day from cultures with moderate density, were placed in replicate vials.…”

Section: Lifespan Assaymentioning

confidence: 99%

“…Locomotion was measured as described by Roshina et al (2014) and Trostnikov et al (2019). Flies were collected and maintained by the same procedures as for the lifespan assays but without recording the deaths.…”

Section: Locomotion Assaymentioning

confidence: 99%

“…For example, the overexpression of the sgg-RB transcript (GenBank 2 # AY122193.1; encodes the major GSK3 isoform (Ruel et al, 1993;Bourouis, 2002), in the nervous system caused pathological changes in neurons paralleled by a rapid decline in survival and severe shortening of lifespan, whereas sgg-RB overexpression in muscles caused only a weak decrease in survival. Overall, in Trostnikov et al (2019), we presented data on the negative effects of strong GSK3 misexpression on lifespan, with the emphasis on the role of the pan-neuronal GSK3 misexpression as a cause of accelerated aging and neuronal pathology.…”

Section: Introductionmentioning

confidence: 99%

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Modulated Expression of the Protein Kinase GSK3 in Motor and Dopaminergic Neurons Increases Female Lifespan in Drosophila melanogaster

et al. 2020

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Section: Introductionmentioning

confidence: 75%

Section: Lifespan Assaymentioning

confidence: 99%

Section: Locomotion Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modulated Expression of the Protein Kinase GSK3 in Motor and Dopaminergic Neurons Increases Female Lifespan in Drosophila melanogaster

et al. 2020

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“…While GSK-3 is a recognized target for the treatment of age related pathologies and multiple diseases, its role in the aging process remains unclear [53]. According to some studies, RNAi knockdown of sgg in Drosophila shortens lifespan or causes lethality [54]. However, these results are contradictory to expectations from earlier studies suggesting that lithium treatment extends lifespan via GSK-3 inhibition, determined using specific RNAi to mediate reduction in sgg expression [55].…”

Section: Lifespan and Locomotor Dysfunction In Sgg Isoa Mutantsmentioning

confidence: 97%

Characterisation of protein isoforms encoded by theDrosophilaGlycogen Synthase Kinase 3 geneshaggy

Korona

Nightingale

Fabre

et al. 2019

Preprint

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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 proteoform classes 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 encoded by independent vertebrate genes. Our analysis suggests that different protein isoforms generated by alternative splicing perform distinct functions.

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Functional analysis of caspase cleavable proteoforms from theDrosophilaGSK-3 geneshaggy

Korona

Nightingale

Fabre

et al. 2020

Preprint

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The Drosophila shaggy (sgg) gene encodes the major fly orthologue of Glycogen Synthase Kinase −3 (GSK-3), a key highly conserved kinase at the heart of many signalling pathways. The sgg locus is complex, encoding multiple protein isoforms that are expressed in distinct temporal and tissue-specific patterns across development. Its isoforms predominantly differ at the carboxy and amino termini due to the use of different transcriptional start sites and alternative splicing events that include internal and terminal exons. One interesting class of proteins isoforms is represented by the Sgg-PD class (Sgg46), three proteoforms that contain a large 582 amino acid N-terminal domain which contains recognition sites for caspase-mediated cleavage. Regulated cleavage at these sites by non-apoptotic caspases has previously been implicated in the regulation of Sgg activity in adult bristle development. Here, we take a genome engineering approach to introduce specific tags into this unique Sgg-PD exon and utilise these for localisation and protein interaction studies. We also generated new loss of function alleles and specific mutations in the caspase cleavage motifs. We find that loss of functions Sgg-PD class alleles are viable and fertile, but exhibit adult locomotor and bristle defects. Expression analysis of lines carrying tags on both sides of the caspase cleavage sites indicates that the cleavage is developmentally regulated during embryogenesis. Surprisingly, we found that in some cells, particularly embryonic hemocytes, the N-terminal domain released by caspase cleavage is retained while the polypeptide containing the conserved kinase domain is apparently lost. Transcriptomic analysis of embryos homozygous for the new caspase-insensitive allele indicates a role for Sgg-PD in the regulation of cytoskeletal and cell junction functions, which is supported by proteomics analysis using specific in locus tags to identify common and unique protein interaction partners with N- and C-terminal domains. Taken together, our work identifies new activities for the Sgg protein and uncovers unexpected roles for caspase cleavage in Sgg biology.

show abstract

Disordered Expression of shaggy, the Drosophila Gene Encoding a Serine-Threonine Protein Kinase GSK3, Affects the Lifespan in a Transcript-, Stage-, and Tissue-Specific Manner

Cited by 12 publications

References 60 publications

Modulated Expression of the Protein Kinase GSK3 in Motor and Dopaminergic Neurons Increases Female Lifespan in Drosophila melanogaster

Modulated Expression of the Protein Kinase GSK3 in Motor and Dopaminergic Neurons Increases Female Lifespan in Drosophila melanogaster

Characterisation of protein isoforms encoded by theDrosophilaGlycogen Synthase Kinase 3 geneshaggy

Functional analysis of caspase cleavable proteoforms from theDrosophilaGSK-3 geneshaggy

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