Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (251K) Freely available
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (6)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Fujimoto, K.
Right arrow Articles by Sugiyama, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fujimoto, K.
Right arrow Articles by Sugiyama, H.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2002, Vol. 30, No. 17 3748-3753
© 2002 Oxford University Press

Sequence-specific protection of plasmid DNA from restriction endonuclease hydrolysis by pyrrole–imidazole–cyclopropapyrroloindole conjugates

Kazuhisa Fujimoto, Hirokazu Iida, Masako Kawakami, Toshikazu Bando, Zhi-Fu Tao and Hiroshi Sugiyama*

Division of Biofunctional Molecules, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Surugadai, Kanda, Chiyoda, Tokyo 101-0062, Japan

*To whom correspondence should be addressed. Tel: +81 03 5280 8032; Fax: +81 03 5280 8127; Email: sugiyama.chem@tmd.ac.jp

The pyrrole–imidazole (Py–Im) triamide–cyclopropa pyrroloindole (CPI) conjugates ImPyImLDu86 (7) and ImImPyLDu86 (14) were synthesized and their alkylating activities and inhibitory effects on DNA hydrolysis by restriction endonucleases were examined. Sequencing gel analysis demonstrated that conjugates 7 and 14 specifically alkylated DNA at 5'-CGCGCG-3' and 5'-PyGGCCPu-3', respectively. Agarose gel electrophoresis indicated that incubation of a supercoiled plasmid, pSPORT I (4109 bp), with conjugate 7 effectively inhibited its hydrolysis by BssHII (5'-G_CGCGC-3'), whereas conjugate 14 had no effect on this hydrolysis. These results suggest that conjugate 7 sequence-specifically inhibits the hydrolysis of DNA by BssHII. Sequence-specific alkylation by the Py–Im triamide–CPI conjugates was further confirmed by inhibition of the Eco52I (5'-C_GGCCG-3') hydrolysis of conjugate 14-treated pQBI PGK (5387 bp). In clear contrast, hydrolysis of pQB1 PGK by DraI (3'-TTT_AAA-3') was not inhibited by 5 µM conjugate 14. That ImImPy did not inhibit the hydrolysis of pQB1 PGK indicates that covalent bond formation is necessary for inhibition. A similar experiment, using linear pQBI PGK, achieved the same extent of protection of the DNA with approximately half the concentration of conjugate 14 as was required to protect supercoiled DNA from hydrolysis.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 HypertensionHome page
E.-H. Yao, N. Fukuda, T. Ueno, H. Matsuda, K. Matsumoto, H. Nagase, Y. Matsumoto, A. Takasaka, K. Serie, H. Sugiyama, et al.
Novel Gene Silencer Pyrrole-Imidazole Polyamide Targeting Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1 Attenuates Restenosis of the Artery After Injury
Hypertension, July 1, 2008; 52(1): 86 - 92.
[Abstract] [Full Text] [PDF]

Home page
 J. Am. Soc. Nephrol.Home page
H. Matsuda, N. Fukuda, T. Ueno, Y. Tahira, H. Ayame, W. Zhang, T. Bando, H. Sugiyama, S. Saito, K. Matsumoto, et al.
Development of Gene Silencing Pyrrole-Imidazole Polyamide Targeting the TGF-beta1 Promoter for Treatment of Progressive Renal Diseases
J. Am. Soc. Nephrol., February 1, 2006; 17(2): 422 - 432.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.