| Project/Area Number |
18590459
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Virology
|
|---|
| Research Institution | National Institute of Infectious Diseases |
|---|
Principal Investigator |
IWAO Kukimoto National Institute of Infectious Diseases, Center for Pathogen Genomics, Lad Head (70291127)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥3,880,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Keywords | human papillomavirus / chromatin / replication / keratinocyte |
|---|
| Research Abstract |
Human papillomavirus type 16 (HPV16) DNA replication, which requires two viral proteins El and E2, occurs only in the differentiating epithelium. Besides the general factors necessary for cellular DNA synthesis, other unidentified cellular factors are supposed to be involved in the regulation of HPV DNA replication. In this study we found that the POU-domain transcription factor hSkn-la, which induces the terminal differentiation of keratinocytes and activates the HPV16 late promoter, enhanced the transient replication of a plasmid containing the HPV16 replication origin in HEK293 cells when co-transfected with a plasmid expressing El and E2. An electrophoretic mobility shift assay with a bacterially expressed hSkn-la or an extract of HeLa cells over-expressing hSkn-la revealed the presence of two hSkn-la binding sites that are distinct from the known three sites, near the replication origin. A chromatin immunoprecipitation analysis showed that hSkn-1 a bound to these sites in cells. Nucleotide substitutions in the sites abolished the binding of hSkn-1 a and the hSkn-1 a-mediated replication enhancement. The data strongly suggest that through the binding to the two sites, hSkn-la enhances 1-IPV DNA replication.
|
