Analysis on alterated chromatin status of viral genome in the host cells.

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670595
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Digestive surgery
• Research Institution: Chiba University
• Principal Investigator: Kaneda Atsushi 千葉大学, 医学(系)研究科(研究院), 教授 (10313024)
• Research Collaborator: MATSUSAKA Keisuke 千葉大学, 医学研究院, 助教 (40610150) ; FUNATA Sayaka 千葉大学, 医学研究院, 特任助教 (80756081) ; OKABE Atsushi 千葉大学, 医学研究院, 特任助教 (80778118)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: エピゲノム

Outline of Final Research Achievements

While virus infection could induce epigenomic alterations in host and viral genomes, the alteration of epigenomic status of viral DNA in host cells is not fully understood. We here challenge clarification and visualization of epigenomic alteration of exogenous viral DNA in host cells. EBV-lacO plasmid DNA with oriP, the EBNA1 gene and lacO repeats, was transfected to 293T cells, and increase of DNA methylation levels was detected at day 50. When GFP-lacR and mCherry-MBD proteins were transfected, these proteins were co-localized in the nucleus, visualizing methylation status of episomal DNA in host cells. Next, a complex formation of methylated EBV-lacO and mCherry-MBD protein was observed under AFM, visualizing methylation status of DNA using AFM. Finally biotinylated EBV-lacO plasmids transfected to 293 cells were collected from the host cells, and the proteins interacting with the episomal DNA were detected by LC/MS/MS analysis, e.g. transcription factors and RNA-binding proteins.

Free Research Field

癌エピゲノム