Functional analysis of CDC7 protein which is interacted with HPV E6.
- Author:
Seo Yun TONG
1
;
Jung Jin KIM
;
Yang Sook CHO
;
Jong Sup PARK
Author Information
1. Department of Obstetrics and Gynecologic, College of Medicine, The Catholic University of Korea, Seoul, Korea. jspark@catholic.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Cervical cancer;
Human papillomavirus;
E6;
Cdc7 kinase
- MeSH:
Carcinogenesis;
Cell Cycle;
Cell Death;
Clone Cells;
Down-Regulation;
Flow Cytometry;
Gene Library;
HeLa Cells;
Humans;
Immunoprecipitation;
Phosphotransferases;
RNA, Small Interfering;
S Phase;
Uterine Cervical Neoplasms;
Yeasts
- From:Korean Journal of Obstetrics and Gynecology
2007;50(3):459-464
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVE: Human papillomaviruses are associated with the majority of cervical cancers. There is a wide effort focused on searching for the target of the involvement of p53-independent HPV-16E6 interacting proteins. We identified Cdc7 (cell division cycle 7-related protein kinases) as a binding partner of E6 and investigated its biological function in cervical cancer cells. METHODS: The baits, E6, fused to the LexA-BD subunit using pBTM116 vector were used to screen an oligoneucleotide primed human HeLa cDNA library cloned in pGAD10 vector. Yeast two-hybrid screens were performed. Immunoprecipitation assay was performed to determine whether E6-Cdc7 interaction occurred. Cells treated with siRNA were analysed by flow cytometry. RESULTS: We have delinated the interactions of E6 with five proteins, namely the IRF-3 (interferon regulatory factor-3), PRKCL?1 (protein kinase C-like 1), PIST (PDZ/coiled-coil domain binding gene), BARD1 (BRCA1 assocated ring domain 1), and Cdc7. The in vitro result of the interaction between E6 and Cdc7 was confirmed by immunoprecipitation experiments. Down-regulation of Cdc7 by small interfering RNA in HeLa cell lines causes an abortive S phase, leading to cell death. CONCLUSION: We have identified the new protein of interaction with HPV E6, Cdc7 kinase. It has been implicated in S phase signaling of cell cycle and the inhibition of Cdc7 induced cell death. But, further investigation is needed to know the biologic function of Cdc7 kinase in cervical carcinogenesis.
