In vivo vizualisation of mono-ADP-ribosylation by dPARP16 upon amino-acid starvation

Aguilera-Gomez, Angelica; Oorschot, Marinke M. van; Veenendaal, Tineke; Rabouille, Cathérine

doi:10.7554/elife.21475

Cited by 47 publications

(79 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Second, their interaction was also suggested in physiological conditions where Rin was observed in very close proximity to Sec16, especially at the earlier time point of amino acid starvation (Figure S3B, S3B’). Third, we used an anchoraway strategy, whereby Sec16 tagged with the Ras CAAX motif resulted in its anchoring to the plasma membrane (Aguilera-Go-mez et al, 2016). Using full-length Sec16-CAAX, we demonstrated that endogenous Rin is very efficiently recruited to the plasma membrane (Figures 3E and 3J), confirming their interaction.…”

Section: Resultsmentioning

confidence: 99%

“…Amino acid starvation is an interesting stress as it triggers the formation of two stress assemblies in the same time frame, both requiring Sec16 but in two different manners: The first, the MARylation of Sec16 on its C terminus by ER-localized dPARP16, is an event that is enough to trigger the formation of Sec bodies (Aguilera-Gomez et al, 2016). The second is the Sec16 interaction and stabilization of phosphoRin, leading to the formation of stress granules.…”

Section: Discussionmentioning

confidence: 99%

“…In this regard, we have recently shown that Sec16 is a key factor driving Sec body formation upon amino acid starvation that activates the ER-localized dPARP16. In turn, dPARP16 mono-ADP-ribosylates Sec16 on a conserved sequence close to its C terminus (Aguilera-Gomez et al, 2016), and Sec16 modification by dPARP16 is enough to elicit Sec body formation. This demonstrates that Sec16 is a stress response protein that plays an important role in the response to amino acid starvation.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Phospho-Rasputin Stabilization by Sec16 Is Required for Stress Granule Formation upon Amino Acid Starvation

et al. 2017

Self Cite

View full text Add to dashboard Cite

SUMMARY Most cellular stresses induce protein translation inhibition and stress granule formation. Here, using Drosophila S2 cells, we investigate the role of G3BP/Rasputin in this process. In contrast to arsenite treatment, where dephosphorylated Ser142 Rasputin is recruited to stress granules, we find that, upon amino acid starvation, only the phosphorylated Ser142 form is recruited. Furthermore, we identify Sec16, a component of the endoplasmic reticulum exit site, as a Rasputin interactor and stabilizer. Sec16 depletion results in Rasputin degradation and inhibition of stress granule formation. However, in the absence of Sec16, pharmacological stabilization of Rasputin is not enough to rescue the assembly of stress granules. This is because Sec16 specifically interacts with phosphorylated Ser142 Rasputin, the form required for stress granule formation upon amino acid starvation. Taken together, these results demonstrate that stress granule formation is fine-tuned by specific signaling cues that are unique to each stress. These results also expand the role of Sec16 as a stress response protein.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Phospho-Rasputin Stabilization by Sec16 Is Required for Stress Granule Formation upon Amino Acid Starvation

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…The regulation of stress granules by ADP-ribosylation is required for the relief of microRNA-mediated translational repression and microRNA-directed mRNA cleavage (Leung et al 2011). A recent study in Drosophila has shown that dPARP-16 and its catalytic activity are required for the formation of Sec body stress assemblies and cell survival in response to amino acid starvation (Aguilera-Gomez et al 2016). Together, these results implicate cytoplasmic PARPs in the post-transcriptional regulation of gene expression.…”

Section: Parps In Rna Biologymentioning

confidence: 95%

“…The discovery of a diverse set of naturally occurring ARBDs, as noted above, has provided new tools for exploring ADP-ribose and PAR in cells through the use of ARBD-GFP fusions, which allow real-time tracking of localized ADP-ribose and PAR synthesis in cells Murawska et al 2011;Aguilera-Gomez et al 2016). The macrodomain from the archaebacterial protein Af1521, which recognizes MAR and the terminal ADP-ribose of PAR, has been fused with a GST tag and used to enrich for ADP-ribosylated targets in genomic and proteomic screens (Bartolomei et al 2016;Martello et al 2016).…”

Section: Adp-ribose Detection Reagentsmentioning

confidence: 99%

PARPs and ADP-ribosylation: recent advances linking molecular functions to biological outcomes

2017

View full text Add to dashboard Cite

The discovery of poly(ADP-ribose) >50 years ago opened a new field, leading the way for the discovery of the poly(ADP-ribose) polymerase (PARP) family of enzymes and the ADP-ribosylation reactions that they catalyze. Although the field was initially focused primarily on the biochemistry and molecular biology of PARP-1 in DNA damage detection and repair, the mechanistic and functional understanding of the role of PARPs in different biological processes has grown considerably of late. This has been accompanied by a shift of focus from enzymology to a search for substrates as well as the first attempts to determine the functional consequences of site-specific ADP-ribosylation on those substrates. Supporting these advances is a host of methodological approaches from chemical biology, proteomics, genomics, cell biology, and genetics that have propelled new discoveries in the field. New findings on the diverse roles of PARPs in chromatin regulation, transcription, RNA biology, and DNA repair have been complemented by recent advances that link ADP-ribosylation to stress responses, metabolism, viral infections, and cancer. These studies have begun to reveal the promising ways in which PARPs may be targeted therapeutically for the treatment of disease. In this review, we discuss these topics and relate them to the future directions of the field.ADP-ribosylation is a reversible post-translational modification (PTM) of proteins resulting in the covalent attachment of a single ADP-ribose unit [i.e., mono(ADP-ribose) (MAR)] or polymers of ADP-ribose units [i.e., poly(ADP-ribose) (PAR)] on a variety of amino acid residues on target proteins (Gibson and Kraus 2012;Daniels et al. 2015a). This modification is mediated by a diverse group of ADPribosyl transferase (ADPRT) enzymes that use ADP-ribose units derived from β-NAD + to catalyze the ADP-ribosylation reaction. These enzymes include bacterial ADPRTs (e.g., cholera toxin and diphtheria toxin) as well as members of three different protein families in yeast and animals: (1) arginine-specific ecto-enzymes (ARTCs), (2) sirtuins, and (3) PAR polymerases (PARPs) . Surprisingly, a recent study showed that the bacterial toxin DarTG can ADP-ribosylate DNA . How this fits into the broader picture of cellular ADP-ribosylation has yet to be determined.In this review, we focus on the mono(ADP-ribosyl)ation (MARylation) and poly(ADP-ribosyl)ation (PARylation) of glutamate, aspartate, and lysine residues by PARP family members. While many reviews have been written on PARPs in the past decade, we highlight the current trends and ideas in the field, in particular those discoveries that have been published in the past 2-3 years.

show abstract

Identification of Ubr1 as an amino acid sensor of steatosis in liver and muscle

Zhao

Zhang

Lin

et al. 2023

J cachexia sarcopenia muscle

View full text Add to dashboard Cite

Background Malnutrition is implicated in human metabolic disorders, including hepatic steatosis and myosteatosis. The corresponding nutrient signals and sensors as well as signalling pathways have not yet been well studied. This study aimed to unravel the nutrient-sensing mechanisms in the pathogenesis of steatosis. Methods Plin2, a lipid droplet (LD) protein-inhibiting lipolysis, is associated with steatosis in liver and muscle. Taking advantage of the Gal4-UAS system, we used the Drosophila melanogaster wing imaginal disc as an in vivo model to study the regulation of Plin2 proteostasis and LD homeostasis. Drosophila Schneider 2 (S2) cells were used for western blotting, immunoprecipitation assays, amino acid-binding assays and ubiquitination assays to further investigate the regulatory mechanisms of Plin2 in response to nutrient signals. Mouse AML12 hepatocytes, human JHH-7 and SNU-475 hepatoma cells were used for immunofluorescence, western blotting and immunoprecipitation to demonstrate that the mode of Plin2 regulation is evolutionarily conserved. In addition, we purified proteins from HEK293 cells and reconstituted in vitro cell-free systems in amino acid-binding assays, pulldown assays and ubiquitination assays to directly demonstrate the molecular mechanism by which Ubr1 senses amino acids to regulate Plin2 proteostasis. Results As a lipolysis inhibitor, Plin2 was significantly elevated in liver (P < 0.05) and muscle (P < 0.05) in patients with steatosis. Consistently, we found that the ubiquitin moiety can be conjugated to any Lys residue in Plin2, ensuring robust clearance of Plin2 by protein degradation. We further demonstrated that the E3 ubiquitin ligase Ubr1 targets Plin2 for degradation in an amino acid-dependent manner. Ubr1 uses two canonical substrate-binding pockets, independent of each other, to bind basic and bulky hydrophobic amino acids, respectively. Mechanistically, amino acid binding allosterically activates Ubr1 by alleviating Ubr1's auto-inhibition. In the absence of amino acids, or when the amino acid-binding capacity of Ubr1 is diminished, Ubr1-mediated Plin2 degradation is inactivated, leading to steatosis. Conclusions We identified Ubr1 as an amino acid sensor regulating Plin2 proteostasis, bridging the knowledge gap between steatosis and nutrient sensing. Our work may provide new strategies for the prevention and treatment of steatosis.

show abstract

In vivo vizualisation of mono-ADP-ribosylation by dPARP16 upon amino-acid starvation

Cited by 47 publications

References 45 publications

Phospho-Rasputin Stabilization by Sec16 Is Required for Stress Granule Formation upon Amino Acid Starvation

Phospho-Rasputin Stabilization by Sec16 Is Required for Stress Granule Formation upon Amino Acid Starvation

PARPs and ADP-ribosylation: recent advances linking molecular functions to biological outcomes

Identification of Ubr1 as an amino acid sensor of steatosis in liver and muscle

Contact Info

Product

Resources

About