Dataset Information

Coronavirus PLPro interactome AP-MS data

ABSTRACT: Main proteases and papain-like proteases (PLpro) are essential coronaviral enzymes required for polypeptide processing during viral maturation. PLpro additionally cleave host cellular proteins to evade anti-viral immune responses. Here, we provide biochemical, structural and functional characterizations of PLpro from SARS-Cov-2 (PLproCoV2) and reveal differences to that of SARS (PLproSARS) in controlling interferon (IFN) and NF-kB pathways. PLproCoV2 and PLproSARS share 89% sequence similarity, yet they differ in their substrate preferences: PLproCoV2 cleaves predominantly ISG15, while PLproSARS targets preferentially ubiquitin chains and Nedd8. The crystal structure of PLproCoV2 in complex with the full-size ISG15 revealed the secondary binding site for the amino-terminal domain of ISG15, thus explaining the affinity and higher specificity, as well as indicating a role for the tyrosine 268 in positioning GRL-0617 inhibitor of PLproCoV2. In human cells, PLproCoV2 cleaves ISG15 from interferon responsive factor 3 (IRF3), blocks its nuclear translocation and reduces type I interferon responses, whereas PLproSARS preferentially mediates deubiquitination and deneddylation of critical components of the NF-kB pathway. Inhibition of PLproCoV2 by GRL-0617 blocks virus-induced cytopathogenic effect, reduces viral release from infected cells and fosters the anti-viral interferon pathway upon infection with SARS-CoV-2. We propose that therapeutic targeting of PLproCoV2 can inhibit SARS-CoV-2 infection and promote anti-viral immunity.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Georg Tascher

LAB HEAD: Ivan Dikic

PROVIDER: PXD018983 | Pride | 2020-08-05

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20200414_GT_AB_TMT16_PLP_test.msf	Msf
	20200414_GT_AB_TMT16_PLP_test.pep.xml	Pepxml
	20200414_GT_AB_TMT16_PLP_test.raw	Raw

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Publications

Papain-like protease regulates SARS-CoV-2 viral spread and innate immunity.

Shin Donghyuk D Mukherjee Rukmini R Grewe Diana D Bojkova Denisa D Baek Kheewoong K Bhattacharya Anshu A Schulz Laura L Widera Marek M Mehdipour Ahmad Reza AR Tascher Georg G Geurink Paul P PP Wilhelm Alexander A van der Heden van Noort Gerbrand J GJ Ovaa Huib H Müller Stefan S Knobeloch Klaus-Peter KP Rajalingam Krishnaraj K Schulman Brenda A BA Cinatl Jindrich J Hummer Gerhard G Ciesek Sandra S Dikic Ivan I

Nature 20200729 7835

The papain-like protease PLpro is an essential coronavirus enzyme that is required for processing viral polyproteins to generate a functional replicase complex and enable viral spread<sup>1,2</sup>. PLpro is also implicated in cleaving proteinaceous post-translational modifications on host proteins as an evasion mechanism against host antiviral immune responses<sup>3-5</sup>. Here we perform biochemical, structural and functional characterization of the severe acute respiratory syndrome coronavi ...[more]

PMID: 32726803

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