Raab, Ulla and Beckenlehner, Karin and Lowin, Tosten and Niller, Hans-Helmut and Doyle, Sean and Modrow, Susanne
(2002)
NS1 Protein of Parvovirus B19 Interacts Directly with DNA Sequences of the p6 Promoter and with the Cellular Transcription Factors Sp1/Sp3.
Virology, 293 (1).
pp. 86-93.
ISSN 0042-6822
Abstract
The nonstructural proteins of parvovirus exert a variety of disparate functions during viral infection ranging from promoter regulation, involvement in DNA replication, and induction of apoptosis. Our interest was focused on the possible mechanism by which the NS1 protein mediates its effects on the p6 promoter of parvovirus B19. It is known that the p6 promoter is highly active in different cell lines and interaction with the viral NS1 protein results in a further increase of the activity. The protein may function by binding directly to the viral DNA or via an indirect binding through interaction with cellular transcription factors bound to the promoter. We examined the interaction of the NS1 protein with cellular transcription factors which are involved in regulating the promoter activity. After purified baculovirus-expressed NS1 protein in gel retardation assays was added, an altered complex formation was observed, indicating that NS1 protein interacts with Sp1/Sp3 transcription factors. Enzyme-linked immunosorbent assays verified these findings. The direct interaction of NS1 protein with p6 promoter elements was analyzed by a coprecipitation assay whereby labeled oligonucleotides spanning the entire promoter region were incubated with NS1 protein followed by an immunoprecipitation with NS1-specific antibodies. An eight-nucleotide-long, almost palindromic sequence (AGGGCGGA) was found as potential NS1-binding motif. Footprint analysis with oligonucleotides containing this DNA motif confirmed this result. Thus, transcriptional regulation by the NS1 protein may involve both the interaction with Sp1/Sp3 that binds to the promoter region and direct binding of NS1 to the promoter DNA.
| Item Type: |
Article
|
| Keywords: |
NS1 Protein; Parvovirus B19; DNA Sequences; p6 Promoter; cellular Transcription Factors Sp1/Sp3; |
| Academic Unit: |
Faculty of Science and Engineering > Biology |
| Item ID: |
7408 |
| Identification Number: |
https://doi.org/10.1006/viro.2001.1285 |
| Depositing User: |
Dr. Sean Doyle
|
| Date Deposited: |
29 Aug 2016 15:28 |
| Journal or Publication Title: |
Virology |
| Publisher: |
Elsevier |
| Refereed: |
Yes |
| URI: |
|
| Use Licence: |
This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available
here |
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