Proteomics analysis of G protein-coupled receptor kinase 4-inhibited cellular growth of HEK293 cells.
J Proteomics
; 207: 103445, 2019 09 15.
Article
em En
| MEDLINE
| ID: mdl-31349021
ABSTRACT
G protein-coupled receptor kinases (GRKs) are involved in a wide range of cellular physiology and pathological activities by specifically phosphorylating activated G protein-coupled receptors (GPCRs) to terminate GPCR signaling, or through regulating non-GPCR substrates. We recently reported that overexpression of GRK4 halts cell proliferation and induces cellular senescent phenotype in HEK293 cells. In this study, a quantitative proteomic assay was performed to analyze the protein profiles between HEK293 cells expressing and not expressing GRK4. Results revealed 39 upregulated and 59 downregulated differently expressed proteins (DEPs) in a total of 4124 identified proteins. Gene ontology (GO) annotation and functional enrichment revealed that the DEPs were related to metabolic processes regulated by the binding of these RNA/proteins under the biological processes. The Kyoto Encyclopedia of Gene and Genomes (KEGG) analysis showed pathways of cell development, division, proliferation, apoptosis, aging, autophagy, cell death and cell cycle progression are involved in. Immunoblotting validation of expression of six key target proteins, CALM1, STAT3, CDK1, CDK6, TOP2A, and GRK4, which speculatively maintain abnormal activity in the above pathways, was consistent with the results of proteomics analysis. Lastly, a biological phenotype assay confirmed that GRK4 promoted HEK293 cell growth blockage and G1/0 arrest. Taken together, this study identified some novel molecules that involve in GRK4 signaling and provided valuable information for further studying the mechanisms underlying GRK4-induced proliferative inhibition. SIGNIFICANCE:
A quantitative proteomic assay was performed in HEK293 cells expressing and not expressing GRK4 39 upregulated and 59 downregulated differently expressed proteins (DEPs)were identified. DEPs involved in pathways of cell development, division, proliferation, apoptosis, aging, autophagy, cell death and cell cycle progression. Biological phenotype assay confirmed that GRK4 prompted HEK293 cell growth blockage and G1/0 arrest.Palavras-chave
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Assunto principal:
Transdução de Sinais
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Regulação da Expressão Gênica
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Fase de Repouso do Ciclo Celular
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Proteômica
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Quinase 4 de Receptor Acoplado a Proteína G
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Pontos de Checagem da Fase G1 do Ciclo Celular
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article