Biology subjects

Leslie Pedrioli, D. M.

Publications and source records attributed to Leslie Pedrioli, D. M..

3 recordsLinked to original sources

bioRxiv2024

PARP14 mediated SQSTM1/p62 cysteine ADP-ribosylation is counteracted by the SARS-CoV-2 macrodomain

Interferon-induced ADP-ribosyltransferases are emerging regulators of cellular innate immunity, yet the relevant substrates and downstream effects remain incompletely defined. Here, we mapped the PARP14-dependent ADP-ribosylome in lung epithelial cancer cells and identified the selective autophagy adaptor SQSTM1/p62, along with multiple innate immune regulators, as IFN-dependent MARylation substrates. Site-resolved analysis revealed that PARP14-dependent MARylation is strongly enriched on cysteine residues, including defined cysteine acceptor sites within p62. PARP14-dependent MARylation is localized to cytoplasmic p62 foci that colocalize with diverse ubiquitin species. Furthermore, our results indicate that TRIM21 limits the autophagic degradation of p62, suggesting that the identified p62 foci may have degradation-independent roles. Consistent with this, both PARP14 and p62 support inflammatory gene expression, linking PARP14-dependent MARylation to innate immune regulation. Together, our findings identify p62 foci as sites of PARP14-dependent ADP-ribosylation and suggest a functional link between interferon signaling, ubiquitin-associated protein assemblies, and MARylation-dependent innate immune regulation.

molecular biology↗

bioRxiv2021

Tankyrase-mediated ADP-ribosylation is a novel regulator of TNF-induced death

Tumor necrosis factor (TNF) is an inflammatory cytokine that, upon binding to its receptor TNFR1, can drive cytokine production, cell survival, or cell death and is a major component of an organisms anti-pathogen repetoire1,2. TNF stimulation leads to the formation of two distinct signalling complexes, a well-defined membrane bound complex (complex 1), and a less well characterised cytosolic death inducing complex (complex 2). Using mass spectrometry, we identified the ADP-ribosyltransferase, tankyrase-1 (TNKS1/TNKS/ARTD5/PARP5a) as a novel native complex 2 component. Following a TNF-induced death stimulus TNKS1 is recruited to complex 2, resulting in complex 2 poly(ADP-ribosyl)ation (PARylation). Tankyrase inhibitors sensitise cells to TNF-induced death, which is correlated with increased complex 2 assembly. Tankyrase-mediated PARylation promotes recruitment of the E3 ligase RNF146 and RNF146 deficiency or proteasome inhibition results in increased levels of complex 2, suggesting that RNF146 causes proteasomal degradation of complex 2. Several viruses express ADP-ribose binding macrodomain proteins, and expression of the SARS-CoV-2 or VEEV macrodomain markedly sensitises cells to TNF-induced death. This suggests that ADP-ribosylation serves as yet another mechanism to detect pathogenic interference of TNF signalling and retaliate with an inflammatory cell death.

cell biology↗

bioRxiv2020

Development of a mass-spectrometry based method for the identification of the in vivo whole blood and plasma ADP-ribosylomes

Blood and plasma proteins are heavily investigated as biomarkers for different diseases. However, the post-translational modification states of these proteins are rarely analyzed since blood contains many enzymes that rapidly remove these modification after sampling. In contrast to the well-described role of protein ADP-ribosylation in cells and organs, its role in blood remains mostly uncharacterized. Here, we discovered that plasma phosphodiesterases and/or ADP-ribosylhydrolases rapidly demodify in vitro ADP-ribosylated proteins. Thus, to identify the in vivo whole blood and plasma ADP-ribosylomes, we established a novel mass-spectrometry based workflow that was applied to blood samples collected from LPS-treated pigs (Sus scrofa), which serves as a model for human systemic inflammatory response syndrome. These analyses identified 60 ADP-ribosylated proteins, 17 of which were ADP-ribosylated plasma proteins. This new protocol provides an important step forward for the rapidly developing field of ADP-ribosylation and defines the blood and plasma ADP-ribosylomes under both healthy and disease conditions.

molecular biology↗