| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Outline of Final Research Achievements |
We identify Wee1, a major regulator at G2/M phase transition, as a cellular factor that interacts with HPV16 E1. Wee1 knockdown reduced the protein levels of exogenously expressed HPV16 E1 in various human cell lines, which resulted from mostly proteasome-independent degradation of E1. Conversely, overexpression of Wee1 increased the level of FLAG-tagged HPV16 E1 in cells. Furthermore, in vitro pull-down assays demonstrated that Wee1 directly bound to HPV16 E1 through the helicase domain, and this domain was required for E1 degradation by Wee1 knockdown. Interestingly, protein levels of the HPV16 E1 ATPase/helicase mutants were not decreased by Wee1 knockdown. Finally, Wee1 knockdown as well as Rad51 knockdown, the main factor of DNA recombination, reduced HPV16 DNA level in W12 cells, which maintain HPV16 genome as episome. Thus, we propose that E1 helicase/ ATPase is stabilized by Wee1 to perform faithful maintenance replication of the viral genome.
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