A unique view of SARS-CoV-2 through the lens of ORF8 protein

Hassan, Sk. Sarif; Ghosh, Shinjini; Attrish, Diksha; Choudhury, Pabitra Pal; Seyran, Murat; Pizzol, Damiano; Adadi, Parise; El-Aziz, Tarek Mohamed Abd; Soares, António; Kandimalla, Ramesh; Lundström, Kenneth; Tambuwala, Murtaza M.; Aljabali, Alaa A. A.; Lal, Amos; Azad, Gajendra Kumar; Uversky, Vladimir N.; Sherchan, Samendra P.; Baetas‐da‐Cruz, Wagner; Uhal, Bruce D.; Rezaei, Nima; Brufsky, Adam

doi:10.1101/2020.08.25.267328

Cited by 15 publications

(20 citation statements)

References 41 publications

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“…We also tested LL-37s' ability to bind to the negatively charged, secreted accessory protein ORF8, which is unique to SARS-CoV-2 and thus linked to efficient pathogen transmission 36,37 . Multiple functions of ORF8 have been proposed including disruption of antigen presentation via down-regulation of MHC I expression and inhibition of interferon I signaling 38,39 .…”

Section: Discussionmentioning

confidence: 99%

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

Roth

Luetke

Meinberger

et al. 2020

Preprint

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Objective: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen accountable for the coronavirus disease 2019 (COVID-19) pandemic. Viral entry via binding of the receptor binding domain (RBD) located within the S1 subunit of the SARS-CoV-2 Spike (S) protein to its target receptor angiotensin converting enzyme (ACE) 2 is a key step in cell infection. The efficient transition of the virus is linked to a unique protein called open reading frame (ORF) 8. As SARS-CoV-2 infections can develop into life threatening lower respiratory syndromes, effective therapy options are urgently needed. Several publications propose vitamin D treatment, although its mode of action against COVID-19 is not fully elucidated. It is speculated that vitamin D's beneficial effects are mediated by up regulating LL-37, a well known antimicrobial peptide with antiviral effects. Methods: Recombinantly expressed SARS-CoV-2 S protein, the extended S1 subunit (S1e), the S2 subunit (S2), the receptor binding domain (RBD), and ORF8 were used for surface plasmon resonance (SPR) studies to investigate LL-37's ability to bind to SARS-CoV-2 proteins and to localize its binding site within the S protein. Binding competition studies were conducted to confirm an inhibitory action of LL-37 on the attachment of SARS-CoV-2 S protein to its entry receptor ACE2. Results: We could show that LL-37 binds to SARS-CoV-2 S protein (LL-37/S-Strep KD = 407 nM, LL-37/S-His KD = 414 nM) with the same affinity, as SARS-CoV-2 binds to hACE2 (hACE2/S-Strep KD = 374 nM, hACE2/S-His KD = 368 nM). The binding is not restricted to the RBD of the S protein, but rather distributed along the entire length of the protein. Interaction between LL-37 and ORF8 was detected with a KD of 294 nM. Further, inhibition of the binding of S-Strep (IC50 = 735 nM), S1e (IC50 = 168 nM), and RBD (IC50 = 126 nM) to hACE2 by LL-37 was demonstrated. Conclusions: We have revealed a biochemical link between vitamin D, LL-37, and COVID-19 severity. SPR analysis demonstrated that LL-37 binds to SARS-CoV-2 S protein and inhibits binding to its receptor hACE2, and most likely viral entry into the cell. This study supports the prophylactic use of vitamin D to induce LL-37 that protects from SARS-CoV-2 infection, and the therapeutic administration of vitamin D for the treatment of COVID-19 patients. Further, our results provide evidence that the direct use of LL-37 by inhalation and systemic application may reduce the severity of COVID-19.

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Section: Discussionmentioning

confidence: 99%

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

Roth

Luetke

Meinberger

et al. 2020

Preprint

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“…Characterization of mutations, especially in fast evolving genes like ORF8 ( Tan et al, 2020 ) is important not only for pathogenesis and immune modulation but also for drugs and diagnostic tests, especially ORF8 which is one of the viral proteins that has been shown to elicit strong and specific antibody response ( Hachim et al, 2020 ; Wang et al, 2020a ). In this context, ORF8 has been studied not only at the evolutionary level ( Pereira, 2020 ; Chen et al, 2020 ) but also at the mutational one ( Hassan et al, 2020 ), in which only 78 mutations in ORF8 were identified.…”

Section: Introductionmentioning

confidence: 99%

Mutational analysis of SARS-CoV-2 ORF8 during six months of COVID-19 pandemic

2021

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“…Among the three mutations (NS8_L84S, NS8_E92K, and NS8_W45L) in NS8 (ORF8) of SARS-CoV-2, the appearance of E92K has not been reported in earlier studies. 44 , 45 …”

Section: Resultsmentioning

confidence: 99%

“… 49 Mutation L84S in ORF8 has been associated with decreased stability of ORF8. 44 L84S destabilizes the folding, which may cause upregulation of the host-immune activity 50 ( Figure 5 ). We predicted the stability effect of MTs NS8_E92K and NS8_W45L through DynaMut online server.…”

Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 Genome from the Khyber Pakhtunkhwa Province of Pakistan

et al. 2021

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Among viral outbreaks, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is one of the deadliest ones, and it has triggered the global COVID-19 pandemic. In Pakistan, until 5th September 2020, a total of 6342 deaths have been reported, of which 1255 were from the Khyber Pakhtunkhwa (KPK) province. To understand the disease progression and control and also to produce vaccines and therapeutic efforts, whole genome sequence analysis is important. In the current investigation, we sequenced a single sample of SARS-CoV-2 genomes (accession no. MT879619) from a male suspect from Peshawar, the KPK capital city, during the first wave of infection. The local SARS-CoV-2 strain shows some unique characteristics compared to neighboring Iranian and Chinese isolates in phylogenetic tree and mutations. The circulating strains of SARS-CoV-2 represent an intermediate evolution from China and Iran. Furthermore, eight complete whole genome sequences, including the current Pakistani isolates which have been submitted to Global Initiative on Sharing All Influenza Data (GSAID), were also investigated for specific mutations and characters. Some novel mutations [NSP2 (D268del), NSP5 (N228K), and NS3 (F105S)] and specific characters have been detected in the coding regions, which may affect viral transmission, epidemiology, and disease severity. The computational modeling revealed that a majority of these mutations may have a stabilizing effect on the viral protein structure. In conclusion, the genome sequencing of local strains is important for better understanding the pathogenicity, immunogenicity, and epidemiology of causative agents.

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A unique view of SARS-CoV-2 through the lens of ORF8 protein

Cited by 15 publications

References 41 publications

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

LL-37 fights SARS-CoV-2: The Vitamin D-Inducible Peptide LL-37 Inhibits Binding of SARS-CoV-2 Spike Protein to its Cellular Receptor Angiotensin Converting Enzyme 2In Vitro

Mutational analysis of SARS-CoV-2 ORF8 during six months of COVID-19 pandemic

SARS-CoV-2 Genome from the Khyber Pakhtunkhwa Province of Pakistan

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