Abstract
The hybridization properties of a series of probes, based on two 9mer oligodeoxynucleotides (designated as I and II) having an appended oligoarginine chain (Rn) to produce peptide-oligonucleotide conjugates or peptide-bridged oligonucleotide pairs (e.g. Rn-I or II-Rn-I), were investigated. For the double-linked probes, we found that the peptide bridge induces the two 9mers to bind complementary single-stranded DNA or RNA targets with substantially enhanced thermal stability. The resulting hybrid with complementary DNA was found to assume a 1:1 complex in the B conformation as judged by UV mixing curves and CD spectroscopy. Complexes of single or double-linked probes with complementary RNA exhibited sensitivity to RNase H digestion. The influence of the identity and chirality of the repeating unit in the bridge, the length of the bridge, the gap size and the salt concentration on the hybridization properties of this new class of oligonucleotide probes was also studied. Our data reveal that these compounds exhibit properties that should prove useful in the development of antisense strategies.
Full Text
The Full Text of this article is available as a PDF (118.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Asseline U., Delarue M., Lancelot G., Toulmé F., Thuong N. T., Montenay-Garestier T., Hélène C. Nucleic acid-binding molecules with high affinity and base sequence specificity: intercalating agents covalently linked to oligodeoxynucleotides. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3297–3301. doi: 10.1073/pnas.81.11.3297. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Breslauer K. J., Frank R., Blöcker H., Marky L. A. Predicting DNA duplex stability from the base sequence. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3746–3750. doi: 10.1073/pnas.83.11.3746. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Calnan B. J., Biancalana S., Hudson D., Frankel A. D. Analysis of arginine-rich peptides from the HIV Tat protein reveals unusual features of RNA-protein recognition. Genes Dev. 1991 Feb;5(2):201–210. doi: 10.1101/gad.5.2.201. [DOI] [PubMed] [Google Scholar]
- Cantor C. R., Warshaw M. M., Shapiro H. Oligonucleotide interactions. 3. Circular dichroism studies of the conformation of deoxyoligonucleotides. Biopolymers. 1970;9(9):1059–1077. doi: 10.1002/bip.1970.360090909. [DOI] [PubMed] [Google Scholar]
- Cazenave C., Chevrier M., Nguyen T. T., Hélène C. Rate of degradation of [alpha]- and [beta]-oligodeoxynucleotides in Xenopus oocytes. Implications for anti-messenger strategies. Nucleic Acids Res. 1987 Dec 23;15(24):10507–10521. doi: 10.1093/nar/15.24.10507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Connell G. J., Illangesekare M., Yarus M. Three small ribooligonucleotides with specific arginine sites. Biochemistry. 1993 Jun 1;32(21):5497–5502. doi: 10.1021/bi00072a002. [DOI] [PubMed] [Google Scholar]
- Durand M., Peloille S., Thuong N. T., Maurizot J. C. Triple-helix formation by an oligonucleotide containing one (dA)12 and two (dT)12 sequences bridged by two hexaethylene glycol chains. Biochemistry. 1992 Sep 29;31(38):9197–9204. doi: 10.1021/bi00153a012. [DOI] [PubMed] [Google Scholar]
- Fita I., Campos J. L., Puigjaner L. C., Subirana J. A. X-ray diffraction study of DNA complexes with arginine peptides and their relation to nucleoprotamine structure. J Mol Biol. 1983 Jun 15;167(1):157–177. doi: 10.1016/s0022-2836(83)80039-4. [DOI] [PubMed] [Google Scholar]
- Furdon P. J., Dominski Z., Kole R. RNase H cleavage of RNA hybridized to oligonucleotides containing methylphosphonate, phosphorothioate and phosphodiester bonds. Nucleic Acids Res. 1989 Nov 25;17(22):9193–9204. doi: 10.1093/nar/17.22.9193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kessler D. J., Pettitt B. M., Cheng Y. K., Smith S. R., Jayaraman K., Vu H. M., Hogan M. E. Triple helix formation at distant sites: hybrid oligonucleotides containing a polymeric linker. Nucleic Acids Res. 1993 Oct 11;21(20):4810–4815. doi: 10.1093/nar/21.20.4810. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leonetti J. P., Rayner B., Lemaitre M., Gagnor C., Milhaud P. G., Imbach J. L., Lebleu B. Antiviral activity of conjugates between poly(L-lysine) and synthetic oligodeoxyribonucleotides. Gene. 1988 Dec 10;72(1-2):323–332. doi: 10.1016/0378-1119(88)90159-x. [DOI] [PubMed] [Google Scholar]
- Olins D. E., Olins A. L., Von Hippel P. H. Model nucleoprotein complexes: studies on the interaction of cationic homopolypeptides with DNA. J Mol Biol. 1967 Mar 14;24(2):157–176. doi: 10.1016/0022-2836(67)90324-5. [DOI] [PubMed] [Google Scholar]
- Ono A., Chen C. N., Kan L. S. DNA triplex formation of oligonucleotide analogues consisting of linker groups and octamer segments that have opposite sugar-phosphate backbone polarities. Biochemistry. 1991 Oct 15;30(41):9914–9912. doi: 10.1021/bi00105a015. [DOI] [PubMed] [Google Scholar]
- Plum G. E., Park Y. W., Singleton S. F., Dervan P. B., Breslauer K. J. Thermodynamic characterization of the stability and the melting behavior of a DNA triplex: a spectroscopic and calorimetric study. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9436–9440. doi: 10.1073/pnas.87.23.9436. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quartin R. S., Brakel C. L., Wetmur J. G. Number and distribution of methylphosphonate linkages in oligodeoxynucleotides affect exo- and endonuclease sensitivity and ability to form RNase H substrates. Nucleic Acids Res. 1989 Sep 25;17(18):7253–7262. doi: 10.1093/nar/17.18.7253. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schatz O., Mous J., Le Grice S. F. HIV-1 RT-associated ribonuclease H displays both endonuclease and 3'----5' exonuclease activity. EMBO J. 1990 Apr;9(4):1171–1176. doi: 10.1002/j.1460-2075.1990.tb08224.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stein C. A., Subasinghe C., Shinozuka K., Cohen J. S. Physicochemical properties of phosphorothioate oligodeoxynucleotides. Nucleic Acids Res. 1988 Apr 25;16(8):3209–3221. doi: 10.1093/nar/16.8.3209. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stevenson M., Iversen P. L. Inhibition of human immunodeficiency virus type 1-mediated cytopathic effects by poly(L-lysine)-conjugated synthetic antisense oligodeoxyribonucleotides. J Gen Virol. 1989 Oct;70(Pt 10):2673–2682. doi: 10.1099/0022-1317-70-10-2673. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sun J. S., François J. C., Montenay-Garestier T., Saison-Behmoaras T., Roig V., Thuong N. T., Hélène C. Sequence-specific intercalating agents: intercalation at specific sequences on duplex DNA via major groove recognition by oligonucleotide-intercalator conjugates. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9198–9202. doi: 10.1073/pnas.86.23.9198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tao J., Frankel A. D. Specific binding of arginine to TAR RNA. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2723–2726. doi: 10.1073/pnas.89.7.2723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thiele D. J., Wang R. W., Leibowitz M. J. Separation and sequence of the 3' termini of M double-stranded RNA from killer yeast. Nucleic Acids Res. 1982 Mar 11;10(5):1661–1678. doi: 10.1093/nar/10.5.1661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tung C. H., Wei Z., Leibowitz M. J., Stein S. Design of peptide-acridine mimics of ribonuclease activity. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7114–7118. doi: 10.1073/pnas.89.15.7114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wei Z., Tung C. H., Zhu T., Stein S. Synthesis of oligoarginine-oligonucleotide conjugates and oligoarginine-bridged oligonucleotide pairs. Bioconjug Chem. 1994 Sep-Oct;5(5):468–474. doi: 10.1021/bc00029a015. [DOI] [PubMed] [Google Scholar]
- Zhu T., Tung C. H., Breslauer K. J., Dickerhof W. A., Stein S. Preparation and physical properties of conjugates of oligodeoxynucleotides with poly(delta)ornithine peptides. Antisense Res Dev. 1993 Winter;3(4):349–356. doi: 10.1089/ard.1993.3.349. [DOI] [PubMed] [Google Scholar]
- Zhu T., Wei Z., Tung C. H., Dickerhof W. A., Breslauer K. J., Georgopoulos D. E., Leibowitz M. J., Stein S. Oligonucleotide-poly-L-ornithine conjugates: binding to complementary DNA and RNA. Antisense Res Dev. 1993 Fall;3(3):265–275. doi: 10.1089/ard.1993.3.265. [DOI] [PubMed] [Google Scholar]
