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C-terminal end of v-src protein interacts with peptide coded by gadd7/adapt15-like RNA in two-hybrid system
O Mizenina1, Y Yanushevich, E Musatkina
1Institute of Carcinogenesis, Cancer Research Center, Moscow, Russia.
Abstract:
The significant differences in the metastatic properties of hamster fibroblasts transformed by the Rous sarcoma virus (RSV) were associated with mutations in the v-src carboxy-terminal region. To identify the capacity of this region for protein-protein interaction the two-hybrid system was used. The cDNA clone (vseap1), producing the protein specifically bound with the v-src C-terminal part in yeast cells in vivo and in GST-fusion system in vitro was isolated. Vseap1 shared 68% of homology with stressful agents induced RNA-gadd7/adapt15. Two vseap1 specific messenger RNAs were identified: 0.9-kbp RNA expressed in all transformed cells and three times less in embryo fibroblasts; 3.1-kbp transcript was deleted in the cells with suppressed v-src activity and H2O2 resistance.
Insights
Researchers identified a protein, vseap1, that interacts with the Rous sarcoma virus (RSV) v-src protein's C-terminal region, impacting cell metastasis. This discovery sheds light on viral oncogenesis and potential therapeutic targets.
Area of Science:
- Molecular Biology
- Virology
- Oncology
Background:
- Metastatic properties of hamster fibroblasts transformed by Rous sarcoma virus (RSV) are linked to mutations in the v-src carboxy-terminal region.
- Understanding protein-protein interactions in this region is crucial for elucidating viral oncogenesis.
Purpose of the Study:
- To identify proteins interacting with the v-src carboxy-terminal region.
- To investigate the role of these interactions in viral transformation and metastasis.
Main Methods:
- Yeast two-hybrid system was employed to screen for interacting proteins.
- In vitro GST-fusion system was used to confirm protein binding.
- RNA analysis was performed to study gene expression patterns.
Main Results:
- A cDNA clone, vseap1, was isolated, encoding a protein that specifically binds to the v-src C-terminal region in both yeast cells (in vivo) and in vitro.
- Vseap1 exhibits 68% homology to RNA-gadd7/adapt15, a gene induced by stressful agents.
- Two vseap1-specific messenger RNAs were identified: a 0.9-kbp RNA present in all transformed cells and a 3.1-kbp transcript absent in cells with suppressed v-src activity and H2O2 resistance.
Conclusions:
- The v-src C-terminal region mediates protein-protein interactions, with vseap1 being a key interacting partner.
- Vseap1 expression and its associated transcripts are linked to v-src activity and cellular resistance to oxidative stress.
- These findings contribute to understanding the molecular mechanisms of RSV-induced transformation and metastasis.