2003 Fiscal Year Final Research Report Summary
Functional analysis of EBNA1 protein by using a recombinant EBV with gene replacement
| Project/Area Number |
14570258
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Virology
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KANDA Teru Hokkaido Univ., Inst. for Genetic Med., Lec., 遺伝子病制御研究所, 助手 (50333472)
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| Project Period (FY) |
2002 – 2003
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| Keywords | EB virus / latent infection / EBNA1 / gene replacement / transcriptional regulation / bacterial artificial chromosome |
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| Research Abstract |
EBNA1 protein, a viral protein encoded by Epstein-Barr virus (EBY), is a multi-functional protein. It is well known to be essential for the replication and segregation of EBV episomes in latently-infected cells, and it also regulates viral gene expression as a transcriptional activator. We generated a recombinant EBV, designated HMG1-EBNA1 knock-in EBV, in which the EBNA1 gene is replaced with a HMGI-EBNA1 chimeilc gene. The HMG1-EBNA1 chimeric gene encodes an EBNA1 mutant with greatly-impaired transcriptional activity, while keeping the ability of EBNA1 to episomally riiaintain EBV based plasmids. Akata cells harboring wild-type EBV and HMG1-EBNA1 knock-in virus were established. The mixture of wild-type EBV and HMGI-EBNA1 knock-in EBV Was used to infect: EBV-negative Akata cells and cord blood lymphocytes. We found that HMG1-EBNA1 knock-in virus was maintained as episomes in Akata cells. By contrast, the analyses of BACmids that had been rescued from the established lymphoblastoid cell lines revealed that the HMG1-EBNA1. chimeric gene was frequently replaced with the wild-type EBNA1 gene, presumably due to the recombination between HMGI-EBNA1 knock-in EBY and wild-type EBV. These results strongly suggest that the regulation of viral gene expression by EBNA1 as a transactivator is critical, for EBV-induced B cell growth transformation.
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Research Products
(3 results)
