Abstract:De-ubiquitylating enzymes (DUBs) reverse protein ubiquitylation and thereby control essential cellular functions. Screening for a DUB that counteracts caspase ubiquitylation to regulate cell survival, we identified the Drosophila ovarian tumour-type DUB DUBA (CG6091). DUBA physically interacts with the initiator caspase death regulator Nedd2-like caspase (Dronc) and de-ubiquitylates it, thereby contributing to efficient inhibitor of apoptosis-antagonist-induced apoptosis in the fly eye. Searching also for non-… Show more
“…Our data also identified transcripts of the Cop9 signalosome genes, CSN5 and CSN6, with strong transcript enrichment in early stages, supporting their known function in early differentiation [ 34 ]. Duba , which has transcript accumulation in the basal region, where elongation takes place, was shown to have an important function in non-apoptotic caspase regulation during late spermatogenesis [ 35 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Metabolic enzymes on the surface of the mitochondria connect CRL3 containing E3 complexes for active degradation of cytoplasmic material during individualisation [ 62 ]. A deubiqutination enzyme, DUBA, is also necessary for individualisation and activation of caspase activity [ 35 ]. This suggests that there is an intimate connection and a fine regulation between mitochondria and the cytosol during the non-apoptotic degradation of cytoplasmic material during individualisation.…”
BackgroundThe formation of matured and individual sperm involves a series of molecular and spectacular morphological changes of the developing cysts in Drosophila melanogaster testis. Recent advances in RNA Sequencing (RNA-Seq) technology help us to understand the complexity of eukaryotic transcriptomes by dissecting different tissues and developmental stages of organisms. To gain a better understanding of cellular differentiation of spermatogenesis, we applied RNA-Seq to analyse the testis-specific transcriptome, including coding and non-coding genes.ResultsWe isolated three different parts of the wild-type testis by dissecting and cutting the different regions: 1.) the apical region, which contains stem cells and developing spermatocytes 2.) the middle region, with enrichment of meiotic cysts 3.) the basal region, which contains elongated post-meiotic cysts with spermatids. Total RNA was isolated from each region and analysed by next-generation sequencing. We collected data from the annotated 17412 Drosophila genes and identified 5381 genes with significant transcript accumulation differences between the regions, representing the main stages of spermatogenesis. We demonstrated for the first time the presence and region specific distribution of 2061 lncRNAs in testis, with 203 significant differences. Using the available modENCODE RNA-Seq data, we determined the tissue specificity indices of Drosophila genes. Combining the indices with our results, we identified genes with region-specific enrichment in testis.ConclusionBy multiple analyses of our results and integrating existing knowledge about Drosophila melanogaster spermatogenesis to our dataset, we were able to describe transcript composition of different regions of Drosophila testis, including several stage-specific transcripts. We present searchable visualizations that can facilitate the identification of new components that play role in the organisation and composition of different stages of spermatogenesis, including the less known, but complex regulation of post-meiotic stages.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-5085-z) contains supplementary material, which is available to authorized users.
“…Our data also identified transcripts of the Cop9 signalosome genes, CSN5 and CSN6, with strong transcript enrichment in early stages, supporting their known function in early differentiation [ 34 ]. Duba , which has transcript accumulation in the basal region, where elongation takes place, was shown to have an important function in non-apoptotic caspase regulation during late spermatogenesis [ 35 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Metabolic enzymes on the surface of the mitochondria connect CRL3 containing E3 complexes for active degradation of cytoplasmic material during individualisation [ 62 ]. A deubiqutination enzyme, DUBA, is also necessary for individualisation and activation of caspase activity [ 35 ]. This suggests that there is an intimate connection and a fine regulation between mitochondria and the cytosol during the non-apoptotic degradation of cytoplasmic material during individualisation.…”
BackgroundThe formation of matured and individual sperm involves a series of molecular and spectacular morphological changes of the developing cysts in Drosophila melanogaster testis. Recent advances in RNA Sequencing (RNA-Seq) technology help us to understand the complexity of eukaryotic transcriptomes by dissecting different tissues and developmental stages of organisms. To gain a better understanding of cellular differentiation of spermatogenesis, we applied RNA-Seq to analyse the testis-specific transcriptome, including coding and non-coding genes.ResultsWe isolated three different parts of the wild-type testis by dissecting and cutting the different regions: 1.) the apical region, which contains stem cells and developing spermatocytes 2.) the middle region, with enrichment of meiotic cysts 3.) the basal region, which contains elongated post-meiotic cysts with spermatids. Total RNA was isolated from each region and analysed by next-generation sequencing. We collected data from the annotated 17412 Drosophila genes and identified 5381 genes with significant transcript accumulation differences between the regions, representing the main stages of spermatogenesis. We demonstrated for the first time the presence and region specific distribution of 2061 lncRNAs in testis, with 203 significant differences. Using the available modENCODE RNA-Seq data, we determined the tissue specificity indices of Drosophila genes. Combining the indices with our results, we identified genes with region-specific enrichment in testis.ConclusionBy multiple analyses of our results and integrating existing knowledge about Drosophila melanogaster spermatogenesis to our dataset, we were able to describe transcript composition of different regions of Drosophila testis, including several stage-specific transcripts. We present searchable visualizations that can facilitate the identification of new components that play role in the organisation and composition of different stages of spermatogenesis, including the less known, but complex regulation of post-meiotic stages.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-5085-z) contains supplementary material, which is available to authorized users.
“…It has also been reported that DUBs can be regulated by allosteric regulation within large multimolecular complexes by phosphorylation, ubiquitination, sumoylation, proteolytic cleavage, and direct stress stimuli, such as reactive oxygen species [ 33 , 55 – 57 ]. Recently, several Drosophila DUBs were shown to be involved in regulation of apoptosis by stabilizing both anti- and proapoptotic cell death machinery proteins [ 38 , 39 , 58 , 59 ]. DUBAI deubiquitinase positively controls the stability of central anti-apoptotic E3 ligase DIAP1.…”
Section: Resultsmentioning
confidence: 99%
“…Another Drosophila DUB which is homologous to human USP36, called scny/et, functions as a direct or indirect positive regulator of DIAP1 [ 38 , 58 ]. The DUBA enzyme (also known as OTUD5), belonging to the OTU class of DUBs, possesses Dronc-controlling activity, whose catalytic activity is positively regulated by phosphorylation [ 39 ]. The present genetic study identified a proapoptotic function for the deubiquitinase DUSP31 , which is involved in the positive regulation of Dark/Dronc apoptosome components.…”
Section: Resultsmentioning
confidence: 99%
“…Recently, it has been shown that deubiquitinase DUBAI stabilizes DIAP1, serving as an additional regulator inhibiting Drosophila apoptosis [ 38 ]. Deubiquitinase DUBA has also been identified as an enzyme associated with regulation of Dronc function [ 39 ]. However, the functions of the majority of predicted DUBs remain unknown.…”
Drosophila have been used to identify new components in apoptosis regulation. The Drosophila protein Dark forms an octameric apoptosome complex that induces the initiator caspase Dronc to trigger the caspase cell death pathway and, therefore, plays an important role in controlling apoptosis. Caspases and Dark are constantly expressed in cells, but their activity is blocked by DIAP1 E3 ligase-mediated ubiquitination and subsequent inactivation or proteasomal degradation. One of the regulatory mechanisms that stabilize proapoptotic factors is the removal of ubiquitin chains by deubiquitinases. In this study performed a modified genetic screen for deubiquitinases (dsRNA lines) to identify those involved in stabilizing proapoptotic components. Loss-of-function alleles of deubiquitinase DUSP31 were identified as suppressors of the Dronc overexpression phenotype. DUSP31 deficiency also suppresses apoptosis induced by the RHG protein, Grim. Genetic analysis revealed for the first time that DUSP31 deficiency sufficiently suppresses the Dark phenotype, indicating its involvement in the control of Dark/Dronc apoptosome function in invertebrate apoptosis.
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