Expression of Drosophila Cabut during early embryogenesis, dorsal closure and nervous system development

Belacortu, Yaiza; Weiss, Ron; Kadener, Sebastián; Paricio, Nuria

doi:10.1016/j.gep.2010.11.004

Cited by 13 publications

(26 citation statements)

References 53 publications

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“…For the repression assay, S2 cells were transfected with 0.5 µg of the pMT-Cbt-V5 plasmid, 0.5 µg of the pHStinger-Prom1-2 plasmid (see [13]) and 0.1 µg of the PAC-cherry vector (to normalize for transfection efficiency) in 6-well plates at 70–90% confluence according to supplier's recommendations (Transfection Custom Insect Reagent, Mirus Bio Corp, Madison, WI). Expression of the recombinant protein was induced by incubation in medium containing 0.25/0.75 µM copper sulfate for 48 h. Fluorescence was measured using a TECAN infinit® 200 plate reader.…”

Section: Methodsmentioning

confidence: 99%

“…After incubation with rabbit anti-β-Gal (1∶1000, Cappel/ICN Biomedicals, Ohio, USA) and mouse monoclonal anti-Lamin (1∶200) (Developmental Studies Hybridoma Bank, Iowa, USA) antibodies for 2 h, cells were washed three times with 0.5% BSA/PBS, incubated with anti-rabbit-FITC (1∶200, Calbiochem EMD Biosciences Inc., La Jolla, CA) and anti-mouse-Cy3 (1∶200, Invitrogen) antibodies for 1 h, washed three times with PBS, mounted in Vectashield (Vector Laboratory, Peterborough, UK) and examined under a Leica TCS SP1 confocal microscope. Ovaries were dissected as described [45] and stained using the anti-CbtΔZn antibody [13]. Scanning electron microscopy analysis of adult eyes was performed following the critical point dry method using a Hitachi S4100 microscope, as previously described [46].…”

Section: Methodsmentioning

confidence: 99%

“…Bruce, personal communication) and cell proliferation and patterning [12]. Experiments in Drosophila embryos and S2 cells have shown that Cbt is a nuclear protein, although it is also present in axons in the central and peripheral nervous systems [13]. Cbt orthologs have been identified in other Drosophila species and insects, including the mosquito ( Anopheles gambiae and Aedes aegypti ), red flour beetle ( Tribolium castaneum ), honeybee ( Apis mellifera ) and silkworm ( Bombyx mori ), as well as in other invertebrate organisms such as ascidians, echinoderms and crustaceans [14], [15].…”

Section: Introductionmentioning

confidence: 99%

“…Cbt orthologs have been identified in other Drosophila species and insects, including the mosquito ( Anopheles gambiae and Aedes aegypti ), red flour beetle ( Tribolium castaneum ), honeybee ( Apis mellifera ) and silkworm ( Bombyx mori ), as well as in other invertebrate organisms such as ascidians, echinoderms and crustaceans [14], [15]. The expression patterns of cbt transcripts and proteins during embryonic development are highly conserved among Drosophilidae [13], [14]. Interestingly, Cbt also presents high similarity to the vertebrate proteins encoded by the TGF-β-inducible early-response genes (TIEGs) [14], [16] and has also been named Drosophila TIEG ( dTIEG ) [12].…”

Section: Introductionmentioning

confidence: 99%

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Transcriptional Activity and Nuclear Localization of Cabut, the Drosophila Ortholog of Vertebrate TGF-β-Inducible Early-Response Gene (TIEG) Proteins

et al. 2012

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BackgroundCabut (Cbt) is a C2H2-class zinc finger transcription factor involved in embryonic dorsal closure, epithelial regeneration and other developmental processes in Drosophila melanogaster. Cbt orthologs have been identified in other Drosophila species and insects as well as in vertebrates. Indeed, Cbt is the Drosophila ortholog of the group of vertebrate proteins encoded by the TGF-ß-inducible early-response genes (TIEGs), which belong to Sp1-like/Krüppel-like family of transcription factors. Several functional domains involved in transcriptional control and subcellular localization have been identified in the vertebrate TIEGs. However, little is known of whether these domains and functions are also conserved in the Cbt protein.Methodology/Principal FindingsTo determine the transcriptional regulatory activity of the Drosophila Cbt protein, we performed Gal4-based luciferase assays in S2 cells and showed that Cbt is a transcriptional repressor and able to regulate its own expression. Truncated forms of Cbt were then generated to identify its functional domains. This analysis revealed a sequence similar to the mSin3A-interacting repressor domain found in vertebrate TIEGs, although located in a different part of the Cbt protein. Using β-Galactosidase and eGFP fusion proteins, we also showed that Cbt contains the bipartite nuclear localization signal (NLS) previously identified in TIEG proteins, although it is non-functional in insect cells. Instead, a monopartite NLS, located at the amino terminus of the protein and conserved across insects, is functional in Drosophila S2 and Spodoptera exigua Sec301 cells. Last but not least, genetic interaction and immunohistochemical assays suggested that Cbt nuclear import is mediated by Importin-α2.Conclusions/SignificanceOur results constitute the first characterization of the molecular mechanisms of Cbt-mediated transcriptional control as well as of Cbt nuclear import, and demonstrate the existence of similarities and differences in both aspects of Cbt function between the insect and the vertebrate TIEG proteins.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transcriptional Activity and Nuclear Localization of Cabut, the Drosophila Ortholog of Vertebrate TGF-β-Inducible Early-Response Gene (TIEG) Proteins

et al. 2012

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“…3). Cbt is known to be involved in the development of the nervous system (Belacortu et al 2011), sox14 is required for 20E signalling at the onset of metamorphosis (Ritter and Beckstead 2010) and sug plays a role in determining adult life span (Landis et al 2003). The TF mbf1 also showed enhanced sensitivity upon ethanol exposure (Fig.…”

Section: Time H Fold Changementioning

confidence: 99%

Functional roles for redox genes in ethanol sensitivity in Drosophila

Awofala

Davies

Jones

2012

Funct Integr Genomics

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Whilst the effects and associated costs of excessive alcohol consumption in the human population are obvious at one level, the roles played by genetic factors at the molecular level are still unclear. Drosophila melanogaster has an alcohol response comparable to humans and is used as a genetic model to study the functional roles of genes regulated in response to ethanol. In the current study, the biological processes associated with behavioural responses to acute alcohol exposure in Drosophila have been analysed using whole genome expression profiling. Ethanol response genes differentially expressed (a) at a single time point (2 h) and (b) in a time series (0-4 h) were identified using microarrays. In addition, a subset of differentially expressed genes was validated using behavioural sedation and recovery assays. The study shows that genes involved in redox processes, neuron development, and specific signalling and metabolic pathways (including glutathione metabolism) form part of the response to ethanol in Drosophila. Biological processes for the regulation of oxidative stress are the common functional denominator of many of the ethanol response genes identified. These upregulated genes work to rescue cells from oxidative stress and its consequences such as protein misfolding, apoptosis and ageing. In the current study, an enrichment of Drosophila genes linked to ageing is observed for the first time. The functional genomics data revealed by such studies can be used to predict transcription networks of ethanol response genes, but the future lies in mapping these networks to the human population, with the ultimate aim of identifying genetic factors for alcohol use disorders.

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Drosophila as a model of wound healing and tissue regeneration in vertebrates

Belacortu

Paricio

2011

Developmental Dynamics

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Understanding the molecular basis of wound healing and regeneration in vertebrates is one of the main challenges in biology and medicine. This understanding will lead to medical advances allowing accelerated tissue repair after wounding, rebuilding new tissues/organs and restoring homeostasis. Drosophila has emerged as a valuable model for studying these processes because the genetic networks and cytoskeletal machinery involved in epithelial movements occurring during embryonic dorsal closure, larval imaginal disc fusion/regeneration, and epithelial repair are similar to those acting during wound healing and regeneration in vertebrates. Recent studies have also focused on the use of Drosophila adult stem cells to maintain tissue homeostasis. Here, we review how Drosophila has contributed to our understanding of these processes, primarily through live-imaging and genetic tools that are impractical in mammals. Furthermore, we highlight future research areas where this insect may provide novel insights and potential therapeutic strategies for wound healing and regeneration. Developmental Dynamics 240:2379-2404,

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Expression of Drosophila Cabut during early embryogenesis, dorsal closure and nervous system development

Cited by 13 publications

References 53 publications

Transcriptional Activity and Nuclear Localization of Cabut, the Drosophila Ortholog of Vertebrate TGF-β-Inducible Early-Response Gene (TIEG) Proteins

Transcriptional Activity and Nuclear Localization of Cabut, the Drosophila Ortholog of Vertebrate TGF-β-Inducible Early-Response Gene (TIEG) Proteins

Functional roles for redox genes in ethanol sensitivity in Drosophila

Drosophila as a model of wound healing and tissue regeneration in vertebrates

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