A Targeted<i>In Vivo</i>RNAi Screen Reveals Deubiquitinases as New Regulators of Notch Signaling

Zhang, Junzheng; Liu, Min; Su, Ying; Du, Juan; Zhu, Alan Jian

doi:10.1534/g3.112.003780

Cited by 38 publications

(37 citation statements)

References 110 publications

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“…In an in vivo RNAi screen targeting Drosophila Ub-proteasome system (UPS) genes (Du et al, 2011;Zhang et al, 2012), we found that inhibiting Ubpy activity resulted in larval wing disc deformation and adult fly lethality, consistent with an essential role of Ubpy in the developing wing (Mukai et al, 2010). Correct patterning of adult wing relies on interplay among several signaling systems, including Hh, Notch (N) and Wingless (Wg) signaling.…”

Section: Resultsmentioning

confidence: 60%

“…In contrast to the lack of effect of Ubpy on Hh signaling, reduced Wg signaling in Ubpy KO clones (supplementary material Fig. S4F; Mukai et al, 2010) and loss of margin bristles caused by Ubpy RNAi (Zhang et al, 2012) were observed. Developmental signaling proteins are trapped in enlarged endosomal vesicles…”

Section: Resultsmentioning

confidence: 77%

“…ap-Gal4 (Du et al, 2011), MS1096-Gal4 (Zhang et al, 2012), ptc-lacZ, dpplacZ (Su et al, 2011), dor 8 (gift of Helmut Krämer) (Sevrioukov et al, 1999), hrs D28 (gift of Hugo Bellen) (Lloyd et al, 2002), Ubpy KO (gift of Satoshi Goto) (Mukai et al, 2010), vps2 PP6 and vps22 ZZ13 (gift of David Bilder) (Vaccari et al, 2009) alleles were described previously. Transgenic RNAi flies targeting different regions of Ubpy gene were obtained from Dr Satoshi Goto (Mitsubishi-Kagaku Institute of Life Sciences, Machida, Japan) (5798R-1 and 5798R-2) and the Vienna Drosophila RNAi Center (VDRC #107623).…”

Section: Fly Geneticsmentioning

confidence: 99%

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Ubpy controls the stability of the ESCRT-0 subunit Hrs in development

Zhang

Lei

et al. 2014

Development

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Ubiquitylated developmental membrane signaling proteins are often internalized for endocytic trafficking, through which endosomal sorting complexes required for transport (ESCRT) act sequentially to deliver internalized cargos to lysosomes. The ESCRT function in endocytic sorting is well established; however, it is not fully understood how the sorting machinery itself is regulated. Here, we show that Ubiquitin isopeptidase Y (Ubpy) plays a conserved role in vivo in the homeostasis of an essential ESCRT-0 complex component Hrs. We find that, in the absence of Drosophila Ubpy, multiple membrane proteins that are essential components of important signaling pathways accumulate in enlarged, aberrant endosomes. We further demonstrate that this phenotype results from endocytic pathway defects. We provide evidence that Ubpy interacts with and deubiquitylates Hrs. In Ubpy-null cells, Hrs becomes ubiquitylated and degraded in lysosomes, thus disrupting the integrity of ESCRT sorting machinery. Lastly, we find that signaling proteins are enriched in enlarged endosomes when Hrs activity is abolished. Together, our data support a model in which Ubpy plays a dual role in both cargo deubiquitylation and the ESCRT-0 stability during development. KEY WORDS: Developmental signaling, Endocytic machinery, ESCRT-0, Hrs, Ubpy, Drosophila INTRODUCTIONMany developmental signaling events are initiated through ligand binding to respective receptors at the plasma membrane. Following uptake into endocytic vesicles referred to as early or sorting endosomes, membrane receptors may recycle back to cell surface via tubular recycling endosomes. However, when directed by a ubiquitin (Ub) signal, receptor complexes are captured and sorted towards lysosomal degradation (Henne et al., 2011). The Ub moieties of protein cargos are recognized by ESCRT complexes, and are sorted into invaginating multivesicular bodies (MVBs). Mature MVBs fuse with lysosomes to deliver protein cargos for degradation (Raiborg and Stenmark, 2009). ESCRT-0 is composed of heterodimers of two subunits: Hrs and Stam. Both subunits bind Ub and clathrin, and Hrs additionally has an FYVE zinc-finger domain that binds phosphatidylinositol 3-phosphate. The ability to bind both lipid and Ub allows ESCRT-0 to initiate endosomal sorting (Clague et al., 2012).Ubpy (or USP8), a USP family deubiquitinase (DUB), participates in sorting of ubiquitylated receptors through its interaction with ESCRT-0 (Mizuno et al., 2005;Row et al., 2006). Most studies of Ubpy focus on endosomal trafficking of growth factor receptor tyrosine kinases (RTKs) in cultured vertebrate cells. However, conflicting data have been reported. In some cases, reduced Ubpy activity results in accumulation of ubiquitylated cargos (Bowers et al., 2006;Mizuno et al., 2006;Row et al., 2006; Alwan and van Leeuwen, 2007). Other studies suggest that Ubpy promotes RTK stability (Mizuno et al., 2005;Niendorf et al., 2007; Berlin et al., 2010). The role of Ubpy in Drosophila development is equally controversial. It was ...

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Section: Resultsmentioning

confidence: 60%

Section: Resultsmentioning

confidence: 77%

Section: Fly Geneticsmentioning

confidence: 99%

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Ubpy controls the stability of the ESCRT-0 subunit Hrs in development

Zhang

Lei

et al. 2014

Development

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“…Deubiquitinases Notch [145] Cholesterol VEGF [146][147][148] MEF2 transcription factors VEGF-Notch [149] Podosomes VEGF-Notch [150] Adipogenic proteins VEGF-Notch [151,152] Glucose regulators VEGF-Notch [152,153] Foxo1 transcription factor EC metabolism [154] Lactate Angiogenesis [155] ROS and redox events VEGF [156][157][158] Cilia Angiogenesis [159,160] It has been shown that EC proliferative capacities can be stimulated by various inducers [41,42,164,165]. A variety of in vivo and in vitro models for understanding EC behaviour during angiogenesis at the cellular level have been derived from systems such as rabbit cornea [166], developing mouse retina [167], intersegmental vessel growth in zebrafish [168] and using ECs embedded in collagen or fibrin gels [169,170].…”

Section: Recently Identified For Sprouting Angiogenesis (Vegf/notch Pmentioning

confidence: 99%

Mechanisms of Angiogenesis Process after Pancreatic Islet Cell Transplantation: Role of Intra-islet Endothelial Cells

Narayanan¹,

Loganathan²,

Dhanasekaran³

et al. 2017

J Transplant Technol Res

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Angiogenic sprouting is a complex, multi-step process involving highly integrated cell behaviours, initial interaction with the environment and signalling pathways. Endothelial cells (ECs) are central to the angiogenic process, with recent insights establishing how these cells communicate with each other and with their microenvironment to form branched vascular networks. Using pancreatic islets as a model for vascularized tissue, this review will present a general overview of EC behaviour dynamics in sprouting angiogenesis, particularly focusing on the interplay between VEGF and Notch pathways. A better understanding of molecular mechanisms associated with intra-islet EC cross-talk and its micro-environment may present exciting new perspectives on islet graft to host revascularization and in supporting islet graft survival.

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“…org/). Of note, the USP encoding gene CG8232 was not included in this study [7,19]. The asterix (*) indicates the Usp genes not screened in [7].…”

Section: Additional Filesmentioning

confidence: 99%

Identifying USPs regulating immune signals in

Engel

Viargues

Mortier

et al. 2014

Cell Communication and Signaling

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Background: Rapid activation of innate immune defences upon microbial infection depends on the evolutionary conserved NF-κB dependent signals which deregulation is frequently associated with chronic inflammation and oncogenesis. These signals are tightly regulated by the linkage of different kinds of ubiquitin moieties on proteins that modify either their activity or their stability. To investigate how ubiquitin specific proteases (USPs) orchestrate immune signal regulation, we created and screened a focused RNA interference library on Drosophila NF-κB-like pathways Toll and Imd in cultured S2 cells, and further analysed the function of selected genes in vivo.

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A TargetedIn VivoRNAi Screen Reveals Deubiquitinases as New Regulators of Notch Signaling

Cited by 38 publications

References 110 publications

Ubpy controls the stability of the ESCRT-0 subunit Hrs in development

Ubpy controls the stability of the ESCRT-0 subunit Hrs in development

Mechanisms of Angiogenesis Process after Pancreatic Islet Cell Transplantation: Role of Intra-islet Endothelial Cells

Identifying USPs regulating immune signals in

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