Engineered variants of the Ras effector protein RASSF5 (NORE1A) promote anticancer activities in lung adenocarcinoma
| Autor: | Hemant Kumar, Yoav Peleg, Ariel Erijman, Julia M. Shifman, Ashish Noronha, Anamika Singh, Yosef Yarden |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
MAPK/ERK pathway
Ras inhibitors Models Molecular Lung Neoplasms Tumor suppressor gene protein-protein interactions SARAH Salvador-RASSF-Hippo Son of Sevenless SPR surface plasmon resonance Adenocarcinoma of Lung GTPase RASSF5 Ras association domain family 5 Biochemistry Nore1A GEF guanine nucleotide exchange factor Ras effector protein PLC phospholipase C APC allophycocyanin ERK extracellular signal-regulated kinase Protein Domains FACS fluorescence-activated cell sorting Humans SOS son of sevenless Genes Tumor Suppressor Molecular Biology Adaptor Proteins Signal Transducing biology Oncogene RBD RAS-binding domain Chemistry Effector protein engineering Cell Biology YSD yeast surface display Cell biology A549 Cells Mitogen-activated protein kinase Mutation biology.protein ras Proteins RASSF5 Guanine nucleotide exchange factor FITC fluorescein isothiocyanate Ras oncogene Apoptosis Regulatory Proteins MEK mitogen-activated protein kinase Research Article Protein Binding |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | Within the superfamily of small GTPases, Ras appears to be the master regulator of such processes as cell cycle progression, cell division, and apoptosis. Several oncogenic Ras mutations at amino acid positions 12, 13, and 61 have been identified that lose their ability to hydrolyze GTP, giving rise to constitutive signaling and eventually development of cancer. While disruption of the Ras/effector interface is an attractive strategy for drug design to prevent this constitutive activity, inhibition of this interaction using small molecules is impractical due to the absence of a cavity to which such molecules could bind. However, proteins and especially natural Ras effectors that bind to the Ras/effector interface with high affinity could disrupt Ras/effector interactions and abolish procancer pathways initiated by Ras oncogene. Using a combination of computational design and in vitro evolution, we engineered high-affinity Ras-binding proteins starting from a natural Ras effector, RASSF5 (NORE1A), which is encoded by a tumor suppressor gene. Unlike previously reported Ras oncogene inhibitors, the proteins we designed not only inhibit Ras-regulated procancer pathways, but also stimulate anticancer pathways initiated by RASSF5. We show that upon introduction into A549 lung carcinoma cells, the engineered RASSF5 mutants decreased cell viability and mobility to a significantly greater extent than WT RASSF5. In addition, these mutant proteins induce cellular senescence by increasing acetylation and decreasing phosphorylation of p53. In conclusion, engineered RASSF5 variants provide an attractive therapeutic strategy able to oppose cancer development by means of inhibiting of procancer pathways and stimulating anticancer processes. |
| Databáze: | OpenAIRE |
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