This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mysiak, M. E.
Right arrow Articles by van der Vliet, P. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mysiak, M. E.
Right arrow Articles by van der Vliet, P. C.

 Previous Article  |  Next Article 

Journal of Virology, February 2004, p. 1928-1935, Vol. 78, No. 4
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.4.1928-1935.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bending of Adenovirus Origin DNA by Nuclear Factor I as Shown by Scanning Force Microscopy Is Required for Optimal DNA Replication

Monika E. Mysiak,1 Marjoleine H. Bleijenberg,1 Claire Wyman,2 P. Elly Holthuizen,1 and Peter C. van der Vliet1*

Department of Physiological Chemistry, University Medical Center Utrecht, and Centre for Biomedical Genetics, 3584 CG Utrecht,1 Department of Radiation Oncology, Erasmus Medical Center-Daniel, and Department of Cell Biology and Genetics, Erasmus Medical Center, 3000 DR Rotterdam, The Netherlands2

Received 25 August 2003/ Accepted 29 October 2003

Nuclear factor I (NFI) is a transcription factor that binds to the adenovirus type 5 (Ad5) origin of replication and recruits the adenovirus DNA polymerase, thereby stimulating initiation of DNA replication in vitro. Using scanning force microscopy, we demonstrate that NFI induces a 60° bend upon binding to the origin. The A/T-rich region preceding the core recognition sequence of NFI influences the DNA bend angle, since substitution of A/T base pairs by G/C base pairs severely decreases bending. Mutations in the A/T-rich region do not affect binding of NFI to DNA. However, mutations that reduce the protein-induced bend lead to a loss of NFI-stimulated replication, indicating that DNA bending is functionally important. In contrast, basal initiation or DNA binding of the polymerase is not impaired by these origin mutations. We conclude that binding of NFI to the Ad5 origin causes structural changes in DNA that are essential for the stimulatory function of NFI in replication. We propose that NFI-induced origin bending facilitates the assembly of a functional initiation complex.

* Corresponding author. Mailing address: UMCU, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands. Phone: (31) 302538989. Fax: (31) 302539035. E-mail: P.C.vanderVliet{at}med.uu.nl.

Journal of Virology, February 2004, p. 1928-1935, Vol. 78, No. 4
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.4.1928-1935.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhang, J., McCauley, M. J., Maher, L. J. III, Williams, M. C., Israeloff, N. E. (2009). Mechanism of DNA flexibility enhancement by HMGB proteins. Nucleic Acids Res 37: 1107-1114 [Abstract] [Full Text]  
  • Dame, R. T., van Mameren, J., Luijsterburg, M. S., Mysiak, M. E., Janicijevic, A., Pazdzior, G., van der Vliet, P. C., Wyman, C., Wuite, G. J.L. (2005). Analysis of scanning force microscopy images of protein-induced DNA bending using simulations. Nucleic Acids Res 33: e68-e68 [Abstract] [Full Text]  
  • Mysiak, M. E., Wyman, C., Holthuizen, P. E., van der Vliet, P. C. (2004). NFI and Oct-1 bend the Ad5 origin in the same direction leading to optimal DNA replication. Nucleic Acids Res 32: 6218-6225 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»