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Chlorocebus sabaeus vero E6 and and eIF4E(S209A) Vero cells (S209A-Vero) LC-MSMS upon infection by coronavirus PEDV

ABSTRACT: Two rounds of TMT relative quantitative proteomics were performed to detect cellular factors involved in p-eIF4E regulation of the synthesis of viral proteins.our first round of screening identified differentially expressed proteins in PEDV-infected cells and mock-infected cells; the cellular pathways involved were mainly the estrogen, cAMP, and calcium signaling pathways. Second round screening identified differentially expressed proteins in the PEDV-infected S209A-Vero cells vs. the PEDV-infected WT-Vero cells; the regulated cellular pathways were found to be mainly in the PI3K-Akt, focal adhesion, and mTOR signaling pathways, and the biological processes and molecular functions in which p-eIF4E played a role were related mainly to metabolism and biogenesis, catalytic activity, and stimuli response.4006 host factors were detected, of which 193 (in brown) were significantly upregulated (ratio ≥1.2, P＜0.05) and 191 (in green) were down-regulated upon PEDV infection (ratio ≤0.83, P＜0.05). 29 of the 191 down-regulated proteins were susceptible to a low level of p-eIF4E . Notably, among the 193 upregulated cellular proteins, 77 were upregulated in the WT-Vero over the S209A-Vero cells , suggesting that the WT-Vero cells are more susceptible to a high level of p-eIF4E.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Cercopithecus Aethiops (green Monkey) (grivet)

TISSUE(S): Epithelial Cell, Kidney

SUBMITTER: jing wang

LAB HEAD: XiaoJia Wang

PROVIDER: PXD033916 | Pride | 2023-03-10

REPOSITORIES: Pride

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Proteomic screening identifies RPLp2 as a specific regulator for the translation of coronavirus.

Dong Hui-Jun HJ Wang Jing J Zhang Xiu-Zhong XZ Li Cui-Cui CC Liu Jian-Feng JF Wang Xiao-Jia XJ

International journal of biological macromolecules 20230109

Viral mRNA of coronavirus translates in an eIF4E-dependent manner, and the phosphorylation of eIF4E can modulate this process, but the role of p-eIF4E in coronavirus infection is not yet entirely evident. p-eIF4E favors the translation of selected mRNAs, specifically the mRNAs that encode proteins associated with cell proliferation, inflammation, the extracellular matrix, and tumor formation and metastasis. In the present work, two rounds of TMT relative quantitative proteomics were used to scre ...[more]

PMID: 36632964

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Project description:MicroRNAs (miRNAs) are small noncoding RNAs that negatively regulate gene expression post-transcriptionally. They play a critical role in developmental and physiological processes and have been implicated in the pathogenesis of several diseases including cancer. To identify miRNA signatures associated with different stages of neoplastic development, we examined the expression profile of 776 primate miRNAs in the following cells: primary African green monkey kidney (pAGMK) cells; spontaneously immortalized, non-tumorigenic, low-passage VERO cells (10-87 LP); tumorigenic, high-passage VERO cells (10-87 HP); and a cell line (10-87 T) derived from a 10-87 HP cell tumor xenograft in athymic nude mice. When compared with pAGMK cells, the majority of miRNAs were expressed at lower levels in 10-87 LP, 10-87 HP, and 10-87 T cells. We identified 10 up-regulated miRNAs whose level of expression correlated with VERO cell evolution from a non-tumorigenic phenotype to a tumorigenic phenotype. Several miRNAs that were components of the tumorigenic phenotype-specific signatures in our AGMK model are also found in a variety of human tumors. This may prove to be of general relevance to the biology of neoplastic development as it occurs both in vivo as well as in vitro. In addition, one or more of these miRNAs could be potential biomarkers for the expression of the tumorigenic phenotype of VERO cells. The spontaneousely transformed VERO cells, non-tumorigenic, were pasasged at low density in culture up to 250. The high passage (p250) was found to be tumorigenic. The cell line from xenograft of high passage was also established. We then evaluated patterns of miRNA expression in pAGMK cells and in derivatives of the 10-87 VERO cell line (10-87 LP cells, 10-87 HP cells, and 10-87 T cells) in an attempt to identify the miRNAs whose altered expression might correlate with, and perhaps be involved in, the evolution of the neoplastic phenotypes that occurred during passage of these AGMK cells in tissue culture. performed high-throughput miRNA profiling to audit the expression level of miRNAs in pAGMK cells and in VERO cells at non-tumorigenic and tumorigenic stages of neoplastic development. The analysis involved pAGMK cells, non-tumorigenic 10-87 low-passage VERO cells (10-87 LP) tumorigenic, high-passage VERO cells (10-87 HP) and a cell line (10-87 T) derived from a 10-87 HP cell tumor xenograft in athymic nude mice.

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