Expression and affinity purification of recombinant mammalian mitochondrial ribosomal small subunit (MRPS) proteins and protein-protein interaction analysis indicate putative role in tumourigenic cellular processes
| Autor: | Jayavelu Subramani, Gopal Gopisetty, Rajkumar Thangarajan, Oviya Revathi Paramasivam |
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| Rok vydání: | 2020 |
| Předmět: |
Ribosomal Proteins
Recombinant Fusion Proteins Breast Neoplasms Mitochondrion Biochemistry Ribosome Chromatography Affinity Protein–protein interaction Mitochondrial Proteins 03 medical and health sciences 0302 clinical medicine Ribosomal protein Gene expression Tumor Cells Cultured Humans Protein Interaction Maps Molecular Biology 030304 developmental biology Glutathione Transferase 0303 health sciences biology Chemistry Translation (biology) General Medicine Fusion protein Glutathione S-transferase 030220 oncology & carcinogenesis biology.protein Female |
| Zdroj: | Journal of biochemistry. 169(6) |
| ISSN: | 1756-2651 |
| Popis: | Mitochondrial ribosomal small subunit (MRPS) group of proteins is structural constituents of the small subunit of mitoribosomes involved in translation. Recent studies indicate role in tumourigenic process, however, unlike cytosolic ribosomal proteins, knowledge on the role of MRPS proteins in alternate cellular processes is very limited. Mapping protein–protein interactions (PPIs) onto known cellular processes can be a valuable tool to identify novel protein functions. In this study, to identify PPIs of MRPS proteins, we have constructed 31 glutathione-S-transferase (GST)/MRPS fusion clones. GST/MRPS fusion proteins were confirmed by MALDI-TOF analysis. GST pull-downs were performed using eight GST/MRPS proteins (MRPS9, MRPS10, MRPS11, MRPS18B, MRPS31, MRPS33, MRPS38 and MRPS39), GST alone as pull-down control and HEK293 cell lysate as the source for anchor proteins followed by nLC/MS/MS analysis and probable PPIs of eight MRPS proteins were identified. Three PPIs from GST pull-downs and interaction between six MRPS proteins and p53 previously reported in PPI database were validated. The PPI network analysis revealed putative role in cellular processes with implications for tumourigenesis. Gene expression screening of a cancer cell line panel indicated overexpression of MRPS10 and MRPS31 in breast cancer. Co-expression module identification tool analysis of breast cancer gene expression and MRPS10 and MRPS31 PPIs revealed putative role for PPI with acyl-CoA dehydrogenase in fatty acid oxidation process regulated by brain-derived neurotrophic factor signalling pathway. |
| Databáze: | OpenAIRE |
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