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Updated: Jul 7, 2026
Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
Published on: February 11, 2019
Screening for novel binding proteins interacting with human papillomavirus type 18 E6 oncogene in the Hela cDNA
1Cancer Biology Center, Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. lee5190008@126.com
Abstract:
To screen for novel binding proteins interacting with high-risk HPV 18 E6 oncogene, the strain AH109 was transformed with pGBKT7-HPV18 E6 plasmid, and subsequent transference was utilized to screen for interacting proteins with HPV 18 E6 in human Hela cDNA library. HPV18 E6 mRNA was expressed in yeast and there was no self-activation and toxicity in AH109. Seven proteins that interacted with HPV18 E6, including transmembrane protein 87B, phosphonoformate immuno-associated protein 5, vimentin, KM-HN-1 protein, dedicator of cytokinesis 7, vaccinia related kinase 2 and a hypothetical protein, were identified. It was suggested that yeast two-hybrid system is an efficient for screening interacting proteins. The high-risk HPV 18 E6 oncogene may interact with the proteins, which may be associated with signal transduction and transcriptional control, epithelial cell invasion and migration, as well as humoral and cellular immune etc. This investigation provides functional clues for further exploration of potential oncogenesis targets for cancer biotherapy.
Insights
Researchers identified seven novel proteins interacting with the high-risk Human Papillomavirus type 18 E6 (HPV 18 E6) oncogene using a yeast two-hybrid system. These interactions offer potential targets for cancer biotherapy.
Area of Science:
- Oncology
- Virology
- Molecular Biology
Background:
- High-risk Human Papillomavirus (HPV) infection is a major cause of cervical cancer.
- The HPV 18 E6 oncogene plays a critical role in viral oncogenesis.
- Identifying HPV 18 E6 interacting proteins is crucial for understanding its carcinogenic mechanisms.
Purpose of the Study:
- To screen for and identify novel cellular proteins that bind to the high-risk HPV 18 E6 oncogene.
- To evaluate the efficacy of the yeast two-hybrid system for identifying protein-protein interactions with HPV 18 E6.
- To provide functional insights into the roles of identified proteins in HPV-associated carcinogenesis.
Main Methods:
- Yeast two-hybrid screening was performed using the AH109 yeast strain.
- The strain was transformed with a pGBKT7-HPV18 E6 plasmid to express the oncogene.
- A human HeLa cDNA library was used to identify interacting proteins.
Main Results:
- The yeast two-hybrid system proved efficient and showed no self-activation or toxicity.
- Seven proteins interacting with HPV 18 E6 were identified: transmembrane protein 87B, phosphonoformate immuno-associated protein 5, vimentin, KM-HN-1 protein, dedicator of cytokinesis 7, vaccinia related kinase 2, and a hypothetical protein.
- These proteins are potentially involved in signal transduction, transcriptional control, cell invasion, migration, and immune responses.
Conclusions:
- The yeast two-hybrid system is an effective method for screening proteins interacting with viral oncogenes.
- The identified proteins interacting with HPV 18 E6 may contribute to oncogenesis through various cellular pathways.
- This study provides functional clues for developing novel cancer biotherapy targets against HPV-associated cancers.
