Ontology highlight
ABSTRACT:
PROVIDER: PRJNA634928 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11848300_1.fastq.gz | Fastqsanger.gz | |||
| SRR11848300_2.fastq.gz | Fastqsanger.gz | |||
| SRR11848301_1.fastq.gz | Fastqsanger.gz | |||
| SRR11848301_2.fastq.gz | Fastqsanger.gz | |||
| SRR11848302_1.fastq.gz | Fastqsanger.gz |
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Project description:Zona pellucida (ZP) proteins, which play an important role in oocyte development, undergo post-translational regulation through furin-mediated cleavage. However, our understanding of the functional significance of furin-mediated cleavage of ZP proteins in female reproduction remains limited. Here, using mouse models with disrupted furin cleavage sites in ZP1, ZP2, and ZP3, we found that loss of the furin site in ZP2 caused female infertility associated with empty follicle syndrome (EFS), characterized by the absence of oocyte retrieval after ovarian hyperstimulation. In contrast, female mice carrying cleavage-resistant variants at the furin sites of ZP1 and ZP3 exhibited defective zona pellucidae in a subset of oocytes, leading to reduced fecundity. Mechanistically, disruption of the furin cleavage site in ZP2 impaired the transmembrane transport of the non-cleaved ZP2 protein and subsequently reduced the levels of SNARE proteins, ultimately triggering oocyte apoptosis through activation of the p53 and PI3K signaling pathways. Collectively, we uncovered the essential role of furin-mediated cleavage of ZP proteins in female fertility and provided new mechanistic insights into the pathogenesis of EFS. These findings open new avenues for investigating the contribution of post-translational modifications to female reproduction and for developing potential therapeutic strategies to treat female infertility.
2025-10-29 | PXD070013 |
Project description:The project aims to determine where recombinant Furin or KLK8 can cleave a motif found at the cleavage site of Activin-A. A mutation at the fifth arginine was also tested in a R310A peptide to look at the effect of this mutation on the cleavage by these two proteases.
2025-02-24 | PXD060918 | Pride
Project description:SARS-CoV-2 USA/WA1 strain stock was sequenced to determine whether the furin cleavage site was intact.
2020-11-28 | GSE162247 | GEO
Project description:The multibasic furin cleavage site at the S1/S2 boundary of the spike protein (S protein) is a hallmark of SARS-CoV-2 and plays an crucial role in viral infection. O-glycosylation near the furin site catalyzed by host cell glycosyltransferases can theoretically hinder spike protein processing and impede viral infection, but so far such a hypothesis has not been tested with authentic viruses. The mechanism underlying furin activation also remains poorly understood. In this study, we have discovered that GalNAc-T3 and T7 jointly initiate clustered O-glycosylations in the multibasic S1/S2 boundary region. These O-glycosylations inhibit furin processing of the spike protein and surprisingly suppress the incorporation of S protein into virus-like-particles (VLPs). Mechanistic analysis revealed that the assembly of spike protein into VLPs relies on protein-protein interaction between the furin-cleaved S protein and a double aspartic motif on the membrane protein of SARS-CoV-2, suggesting a novel mechanism for furin activation of the S protein. Interestingly, a point mutation at P681, found in the alpha and delta variants of SARS-CoV-2, confers resistance to glycosylation by GalNAc-T3 and T7, thereby diminishing the inhibitory effect against furin processing. However, an additional mutation at N679 in the most recent omicron variant reverses this resistance, rendering it susceptible to glycosylation in vitro and sensitive to the expression of GalNAc-T3 and T7 in human lung cells. Together, our findings suggest a glycosylation-based defense mechanism employed by host cells against SARS-CoV-2 and unveil the intricate interplay between the host and pathogen at this critical “battlefield” as the virus initially evades and currently succumbs to host cell glycosylation..
2024-03-21 | PXD049110 | Pride
Project description:Experimental golden hamsters were infected with BANAL-20-236, WK-521 (a parental strain of SARS-CoV-2), and WK-521 dFCS (a WK-521 without furin cleavage site [FCS]) to investigate virological properties and host response in vivo.
2024-05-20 | GSE239893 | GEO
Project description:Severe cardiovascular complications can occur in coronavirus disease of 2019 (COVID-19) patients. Cardiac damage is attributed mostly to the aberrant host response to acute respiratory infection. However, direct infection of cardiac tissue by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) also occurs. We examined here the cardiac tropism of SARS-CoV-2 in spontaneously beating human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).These cardiomyocytes express the angiotensin-converting enzyme 2 (ACE2) receptor but not the transmembrane protease serine 2 (TMPRSS2) that mediates spike protein cleavage in the lungs. Nevertheless, SARS-CoV-2 infection of hiPSC-CMs was prolific: viral transcripts accounted for about 88% of total mRNA. In the cytoplasm of infected hiPSC33 CMs, smooth walled exocytic vesicles contained numerous 65-90 nm particles with canonical ribonucleocapsid structures, and virus-like particles with knob-like spikes covered the cell surface. To better understand how SARS-CoV-2 spreads in hiPSC-CMs we engineered an expression vector coding for the spike protein with a monomeric emerald-green fluorescent protein fused to its cytoplasmic tail (S-mEm). Proteolytic processing of S-mEm and the parental spike were equivalent. Live cell imaging tracked spread of S-mEm cell-to-cell and documented formation of syncytia. A cell-permeable, peptide-based molecule that blocks the catalytic site of furin and furin-like proteases abolished cell fusion. A spike mutant with the single amino acid change R682S that disrupts the multibasic furin cleavage motif was fusion inactive. Thus, SARS42 CoV-2 replicates efficiently in hiPSC-CMs and furin and/or furin-like-protease activation of its spike protein is required for fusion-based cytopathology. This hiPSC-CM platform enables target-based drug discovery in cardiac COVID-19.
2021-09-27 | GSE184715 | GEO
Project description:3 strains of SARS-CoV-2 were created to the function of the QTQTN motif adjacent to the Furin cleavage site on Spike protein: WT (QTQTN), Mutant 1 (QTQVN), Mutant 2 (QVQVN).
2022-07-20 | PXD034332 | Pride
Project description:Addressing the elusive specificity of cysteine cathepsins, which in contrast to caspases and trypsin-like proteases lack strict specificity determining P1 pocket, was calling for innovative approaches. Proteomic analysis of cell lysates with human cathepsins K, V, B, L, S, and F identified 30,000 cleavage sites, which were analyzed by software platform SAPS-ESI (Statistical Approach to Peptidyl Substrate-Enzyme Specific Interactions). SAPS-ESI was used to generate clusters and training sets for support vector machine learning. Cleavage site predictions on the SARS-CoV-2 S protein, confirmed experimentally, exposed the most probable first cut under physiological conditions and suggested furin-like behavior of cathepsins. Crystal structure analysis of representative peptides in complex with cathepsin V revealed rigid and flexible sites consistent with analysis of proteomics data by SAPS-ESI that correspond to positions with heterogeneous and homogeneous distribution of residues. Thereby support for design of selective cleavable linkers of drug conjugates and drug discovery studies is provided.
2023-03-29 | PXD041185 | Pride
Project description:SARS-CoV-2 has caused the largest known coronavirus pandemic and is believed to have emerged from insectivorous bats. Little is known about the evolution of these viruses in their reservoir bat species. In this study, we investigated SARS-CoV-2-host interaction using human and bat cells. Bat cells mount a robust and early antiviral response but elicit a dampened pro-inflammatory response upon SARS-CoV-2 infection compared to human cells. Furthermore, an inactivating R685P mutation within the furin cleavage site (FCS) of the SARS-CoV-2 spike protein was naturally selected for in infected bat cells. Taken together, our data demonstrate that insectivorous bat cells have evolved a differential antiviral immune response against SARS-CoV-2 infection, likely to mitigate immunopathology that is observed in humans. Our study sheds light on the evolution of sarbecoviruses in bats and extends molecular evidence to data from field studies that have demonstrated that SARS-CoV-2-related viruses in wild-caught bats lack an intact FCS.
2025-06-04 | GSE285756 | GEO
Project description:Direct RNA sequencing using an Oxford Nanopore MinION characterised the transcriptome of SARS-CoV-2 grown in Vero E6 cells. This cell line is being widely used to propagate the novel coronavirus. The viral transcriptome was analysed using a recently developed ORF-centric pipeline. This revealed the pattern of viral transcripts, (i.e. subgenomic mRNAs), generally fitted the predicted replication and transcription model for coronaviruses. A 24 nt in-frame deletion was detected in subgenomic mRNAs encoding the spike (S) glycoprotein. This feature was identified in over half of the mapped transcripts and was predicted to remove a proposed furin cleavage site from the S glycoprotein. This motif directs cleavage of the S glycoprotein into functional subunits during virus entry or exit. Cleavage of the S glycoprotein can be a barrier to zoonotic coronavirus transmission and affect viral pathogenicity. Allied to this transcriptome analysis, tandem mass spectrometry was used to identify over 500 viral peptides and 44 phosphopeptides, covering almost all of the proteins predicted to be encoded by the SARS-CoV-2 genome, including peptides unique to the deleted variant of the S glycoprotein. Detection of an apparently viable deletion in the furin cleavage site of the S glycoprotein reinforces the point that this and other regions of SARS-CoV-2 proteins may readily mutate. This is of clear significance given the interest in the S glycoprotein as a potential vaccine target and the observation that the furin cleavage site likely contributes strongly to the pathogenesis and zoonosis of this virus. The viral genome sequence should be carefully monitored during the growth of viral stocks for research, animal challenge models and, potentially, in clinical samples. Such variations may result in different levels of virulence, morbidity and mortality.
2020-03-31 | PXD018241 | Pride