RNAi screening reveals a large signaling network controlling the Golgi apparatus in human cells

Chia, Joanne; Goh, Germaine Yen Lin; Racine, Victor; Ng, Shi Ling; Kumar, Pankaj; Bard, Frédéric

doi:10.1038/msb.2012.59

Cited by 123 publications

(116 citation statements)

References 104 publications

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“…Several siRNA-based screens have been used to study endocytosis, ER to Golgi transport or post-Golgi transport to the plasma membrane (Chia et al, 2012;Collinet et Simpson et al, 2012). Among the genes identified in our screen, 47 have been shown to affect clathrin-dependent endocytosis (Collinet et al, 2010) (supplementary material Table S4), consistent with the notion that MPRs are endocytosed through clathrin-coated pits.…”

Section: Discussionsupporting

confidence: 63%

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A high throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking

Anitei

Ramu

Czupalla

et al. 2014

Journal of Cell Science

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The delivery of newly synthesized soluble lysosomal hydrolases to the endosomal system is essential for lysosome function and cell homeostasis. This process relies on the proper trafficking of the mannose 6-phosphate receptors (MPRs) between the trans-Golgi network (TGN), endosomes and the plasma membrane. Many transmembrane proteins regulating diverse biological processes ranging from virus production to the development of multicellular organisms also use these pathways. To explore how cell signaling modulates MPR trafficking, we used high-throughput RNA interference (RNAi) to target the human kinome and phosphatome. Using high-content image analysis, we identified 127 kinases and phosphatases belonging to different signaling networks that regulate MPR trafficking and/or the dynamic states of the subcellular compartments encountered by the MPRs. Our analysis maps the MPR trafficking pathways based on enzymes regulating phosphatidylinositol phosphate metabolism. Furthermore, it reveals how cell signaling controls the biogenesis of post-Golgi tubular carriers destined to enter the endosomal system through a SRCdependent pathway regulating ARF1 and RAC1 signaling and myosin II activity.

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Section: Discussionsupporting

confidence: 63%

“…4A). These included the dualspecificity phosphatase DUSP2 (dual specificity phosphatase 2), shown to control Golgi integrity (Chia et al, 2012), as well as kinases participating in the p38 and SAPK/JNK signaling cascades (Fig. 4E).…”

Section: Mapk Signaling Pathwaysmentioning

confidence: 99%

A high throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking

Anitei

Ramu

Czupalla

et al. 2014

Journal of Cell Science

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“…Our findings demonstrate that changes in the number of ERES are part of the cellular response to mitogen treatment and therefore imply a close link between the regulation of secretion and cell growth and proliferation. An integrative view on the secretory pathway becomes even more important when aiming at deconvolving the results of recent systems biology approaches that have identified numerous signaling, metabolic or transcriptional regulators that have an effect on the secretory pathway at various levels (Bard et al, 2006;Farhan et al, 2010;Kondylis et al, 2011;Simpson et al, 2012;Chia et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Regulation of Sec16 levels and dynamics links proliferation and secretion

Tillmann

Reiterer

Baschieri

et al. 2014

Journal of Cell Science

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We currently lack a broader mechanistic understanding of the integration of the early secretory pathway with other homeostatic processes such as cell growth. Here, we explore the possibility that Sec16A, a major constituent of endoplasmic reticulum exit sites (ERES), acts as an integrator of growth factor signaling. Surprisingly, we find that Sec16A is a short-lived protein that is regulated by growth factors in a manner dependent on Egr family transcription factors. We hypothesize that Sec16A acts as a central node in a coherent feed-forward loop that detects persistent growth factor stimuli to increase ERES number. Consistent with this notion, Sec16A is also regulated by short-term growth factor treatment that leads to increased turnover of Sec16A at ERES. Finally, we demonstrate that Sec16A depletion reduces proliferation, whereas its overexpression increases proliferation. Together with our finding that growth factors regulate Sec16A levels and its dynamics on ERES, we propose that this protein acts as an integrator linking growth factor signaling and secretion. This provides a mechanistic basis for the previously proposed link between secretion and proliferation.

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“…Furthermore, a recent study from a medium scale RNAi screen identified human sybl1 (encoding for VAMP7) as one of the hits leading to cis Golgi compactness defect. 36 Therefore, we sought to assay for a structural and/or dynamic defect of the Golgi in the absence of VAMP7 expression. To this aim, WT and VAMP7 KO MEFs transiently expressing GFP-tagged N-acetyl-glucosaminyl-transferase (NAGT-GFP) were treated with BFA, a drug known to reversibly disrupt the Golgi apparatus and causes resident enzymes such as NAGT-GFP, to diffuse back to the ER 37 (Fig.…”

Section: Loss Of Vamp7 Affects Golgi Dynamicsmentioning

confidence: 99%

Role of tetanus neurotoxin insensitive vesicle-associated membrane protein in membrane domains transport and homeostasis

et al. 2015

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Keywords: exocytosis, Golgi apparatus, SNARE, sphingolipids, TI-VAMP/VAMP7 Abbreviations: BFA, Brefeldin A; Cer, Ceramide; ER, Endoplasmic Reticulum; GlcCer, Glucosylceramide; GM3, ganglioside monosialic acid 3; GPI, Glycosylphosphatidylinositol; GSL, Glycosphingolipids; LC, Long Chain; PI, Phosphatidylinositide; PM, Plasma Membrane; SM, Sphingomyelin; TGN, D Trans-Golgi Network; TI-VAMP/VAMP7, Tetanus neurotoxin-insensitive vesicle-associated membrane protein / Vesicle associated membrane protein 7; VLC, very long vhain; VSVG, Vesicular Stomatitis Virus GlycoproteinBiological membranes in eukaryotes contain a large variety of proteins and lipids often distributed in domains in plasma membrane and endomembranes. Molecular mechanisms responsible for the transport and the organization of these membrane domains along the secretory pathway still remain elusive. Here we show that vesicular SNARE TI-VAMP/VAMP7 plays a major role in membrane domains composition and transport. We found that the transport of exogenous and endogenous GPI-anchored proteins was altered in fibroblasts isolated from VAMP7-knockout mice. Furthermore, disassembly and reformation of the Golgi apparatus induced by Brefeldin A treatment and washout were impaired in VAMP7-depleted cells, suggesting that loss of VAMP7 expression alters biochemical properties and dynamics of the Golgi apparatus. In addition, lipid profiles from these knockout cells indicated a defect in glycosphingolipids homeostasis. We conclude that VAMP7 is required for effective transport of GPI-anchored proteins to cell surface and that VAMP7-dependent transport contributes to both sphingolipids and Golgi homeostasis.

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RNAi screening reveals a large signaling network controlling the Golgi apparatus in human cells

Cited by 123 publications

References 104 publications

A high throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking

A high throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking

Regulation of Sec16 levels and dynamics links proliferation and secretion

Role of tetanus neurotoxin insensitive vesicle-associated membrane protein in membrane domains transport and homeostasis

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