In Vivo RNAi Screen Reveals Neddylation Genes as Novel Regulators of Hedgehog Signaling

Du, Juan; Zhang, Junzheng; Su, Ying; Liu, Min; Ospina, Jason K.; Yang, Shengyuan; Zhu, Alan Jian

doi:10.1371/journal.pone.0024168

Cited by 23 publications

(25 citation statements)

References 90 publications

(150 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To determine the TfR levels at the cell surface, we preincubated cells with 50 μg/ml of cychloheximide (CHX) for 2 h [23], which inhibits protein synthesis to eliminate possible complications caused by α‐syn effects on TfR synthesis, and the total internal reflection fluorescence microscopy (TIRFM) was used to selectively visualize the cell surface TfR for each condition (Fig. 5 A).…”

Section: Resultsmentioning

confidence: 99%

The secreted oligomeric form of α‐synuclein affects multiple steps of membrane trafficking

et al. 2013

View full text Add to dashboard Cite

a b s t r a c t a-Synuclein (a-syn) can be secreted from neurons into the extracellular space, affecting the homeostasis of neighboring cells, but the pathophysiology of secreted a-syn remains largely unknown. We found that when exogenously applied to COS-7 cells, a-syn secreted from differentiated SH-SY5Y cells was taken up by dynamin-dependent endocytosis. Upon internalization, a-syn significantly increased the rate of transferrin receptor (TfR) internalization and recycling, and subsequently the surface levels of TfR. The effects are attributable to the oligomeric form, but not monomeric or fibrillar form, of extracellular a-syn. Together, multiple alterations in membrane trafficking by secreted oligomeric a-syn may contribute to the early stages of pathogenesis in Parkinson's disease.

show abstract

Section: Resultsmentioning

confidence: 99%

The secreted oligomeric form of α‐synuclein affects multiple steps of membrane trafficking

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Act5C -Gal4, Act5C > yw > Gal4, C96 -Gal4, dpp -Gal4, GMR -Gal4, ptc -Gal4, tub -Gal4, Gbe + Su(H)m8-lacZ [ Su(H)-lacZ ], and neuralized A101 -lacZ ( neur-lacZ ) were described previously (Huang et al 1991; Furriols and Bray 2001; Ahmed et al 2003; Kim et al 2006; Du et al 2011; Su et al 2011). Transgenic RNAi flies targeting all 45 DUB genes predicted in the fly genome were obtained from the Vienna Drosophila RNAi Center [VDRC, Vienna, Austria (Dietzl et al 2007)], and the Fly Stocks of National Institute of Genetics (NIG-Fly; National Institute of Genetics, Shizuoka, Japan; Table S2).…”

Section: Methodsmentioning

confidence: 99%

“…Our primary RNAi screens were conducted at 29° by crossing individual RNAi lines with the dpp -Gal4 and c96 -Gal4 drivers, respectively, for defects on patterning the adult wing margin and scutellar bristles. Crosses were shifted to lower temperatures (18° or 21°) for RNAi lines whose overexpression resulted in embryonic or larval lethality at 29° (Du et al 2011). …”

Section: Methodsmentioning

confidence: 99%

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

Zhang¹,

Liu²,

et al. 2012

G3 Genes|Genomes|Genetics

Self Cite

View full text Add to dashboard Cite

Notch signaling is highly conserved in all metazoan animals and plays critical roles in cell fate specification, cell proliferation, apoptosis, and stem cell maintenance. Although core components of the Notch signaling cascade have been identified, many gaps in the understanding of the Notch signaling pathway remain to be filled. One form of posttranslational regulation, which is controlled by the ubiquitin-proteasome system, is known to modulate Notch signaling. The ubiquitination pathway is a highly coordinated process in which the ubiquitin moiety is either conjugated to or removed from target proteins by opposing E3 ubiquitin ligases and deubiquitinases (DUBs). Several E3 ubiquitin ligases have been implicated in ubiquitin conjugation to the receptors and the ligands of the Notch signaling cascade. In contrast, little is known about a direct role of DUBs in Notch signaling in vivo. Here, we report an in vivo RNA interference screen in Drosophila melanogaster targeting all 45 DUBs that we annotated in the fly genome. We show that at least four DUBs function specifically in the formation of the fly wing margin and/or the specification of the scutellar sensory organ precursors, two processes that are strictly dependent on the balanced Notch signaling activity. Furthermore, we provide genetic evidence suggesting that these DUBs are necessary to positively modulate Notch signaling activity. Our study reveals a conserved molecular mechanism by which protein deubiquitination process contributes to the complex posttranslational regulation of Notch signaling in vivo.

show abstract

“…Elevated de-ubiquitination in the dorsal compartment of the wing disc protected Cubitus interruptus (Ci) protein ( Figure 5D; cf. Figure S7C), a known ubiquitinated substrate subject to proteasomal degradation (Du et al, 2011), from degradation, but not Pc protein ( Figure 5E). These findings together indicate that Pc may be degraded through a Ub-independent mechanism in the proteasome (Erales and Coffino, 2014).…”

Section: The Ubl Domain Of Stx Is Required For Regulated Pc Degradationmentioning

confidence: 99%

Stuxnet Facilitates the Degradation of Polycomb Protein during Development

Zhang

et al. 2016

Developmental Cell

Self Cite

View full text Add to dashboard Cite

Polycomb-group (PcG) proteins function to ensure correct deployment of developmental programs by epigenetically repressing target gene expression. Despite the importance, few studies have been focused on the regulation of PcG activity itself. Here, we report a Drosophila gene, stuxnet (stx), that controls Pc protein stability. We find that heightened stx activity leads to homeotic transformation, reduced Pc activity, and de-repression of PcG targets. Conversely, stx mutants, which can be rescued by decreased Pc expression, display developmental defects resembling hyperactivation of Pc. Our biochemical analyses provide a mechanistic basis for the interaction between stx and Pc; Stx facilitates Pc degradation in the proteasome, independent of ubiquitin modification. Furthermore, this mode of regulation is conserved in vertebrates. Mouse stx promotes degradation of Cbx4, an orthologous Pc protein, in vertebrate cells and induces homeotic transformation in Drosophila. Our results highlight an evolutionarily conserved mechanism of regulated protein degradation on PcG homeostasis and epigenetic activity.

show abstract

In Vivo RNAi Screen Reveals Neddylation Genes as Novel Regulators of Hedgehog Signaling

Cited by 23 publications

References 90 publications

The secreted oligomeric form of α‐synuclein affects multiple steps of membrane trafficking

The secreted oligomeric form of α‐synuclein affects multiple steps of membrane trafficking

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

Stuxnet Facilitates the Degradation of Polycomb Protein during Development

Contact Info

Product

Resources

About