Abstract
The three-dimensional structure of a short DNA fragment, d(GCGAAGC) exhibiting an extraordinarily stable hairpin structure was determined by nuclear magnetic resonance spectroscopy. Two possible models were obtained by molecular modelling using distance and torsion constraints. Only one of these two models is the correct structure, which can clearly explain all the 1H chemical shifts. d(GCGAAGC) is folded back on itself between A4 and A5, and all the sugars in the fragment adopt the C2'-endo conformation. This compact molecule is stabilized by regular extensive base-stacking interaction within each B-form helical strand of G1C2G3A4 and A5G6C7, and by two G-C and one G3-A5 base pairs. The molecule is hard to differentiate into stem and loop regions, so that we classify it as a turn (hairpin-turn) structure experted by a single-stranded DNA. This highly stacked structure shows high thermostability and strong resistance against nucleases contained in E. coli extracts and in human serum.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adhin M. R., Alblas J., van Duin J. Secondary structure at the 3' terminal region of RNA coliphages: comparison with tRNA. Biochim Biophys Acta. 1990 Aug 27;1050(1-3):110–118. doi: 10.1016/0167-4781(90)90150-z. [DOI] [PubMed] [Google Scholar]
- Antao V. P., Lai S. Y., Tinoco I., Jr A thermodynamic study of unusually stable RNA and DNA hairpins. Nucleic Acids Res. 1991 Nov 11;19(21):5901–5905. doi: 10.1093/nar/19.21.5901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Antao V. P., Tinoco I., Jr Thermodynamic parameters for loop formation in RNA and DNA hairpin tetraloops. Nucleic Acids Res. 1992 Feb 25;20(4):819–824. doi: 10.1093/nar/20.4.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Arai K., Low R., Kobori J., Shlomai J., Kornberg A. Mechanism of dnaB protein action. V. Association of dnaB protein, protein n', and other repriming proteins in the primosome of DNA replication. J Biol Chem. 1981 May 25;256(10):5273–5280. [PubMed] [Google Scholar]
- Bianchi M. E. Interaction of a protein from rat liver nuclei with cruciform DNA. EMBO J. 1988 Mar;7(3):843–849. doi: 10.1002/j.1460-2075.1988.tb02883.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boulard Y., Gabarro-Arpa J., Cognet J. A., Le Bret M., Guy A., Téoule R., Guschlbauer W., Fazakerley G. V. The solution structure of a DNA hairpin containing a loop of three thymidines determined by nuclear magnetic resonance and molecular mechanics. Nucleic Acids Res. 1991 Oct 11;19(19):5159–5167. doi: 10.1093/nar/19.19.5159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheong C., Varani G., Tinoco I., Jr Solution structure of an unusually stable RNA hairpin, 5'GGAC(UUCG)GUCC. Nature. 1990 Aug 16;346(6285):680–682. doi: 10.1038/346680a0. [DOI] [PubMed] [Google Scholar]
- Cowing D. W., Bardwell J. C., Craig E. A., Woolford C., Hendrix R. W., Gross C. A. Consensus sequence for Escherichia coli heat shock gene promoters. Proc Natl Acad Sci U S A. 1985 May;82(9):2679–2683. doi: 10.1073/pnas.82.9.2679. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davis P. W., Thurmes W., Tinoco I., Jr Structure of a small RNA hairpin. Nucleic Acids Res. 1993 Feb 11;21(3):537–545. doi: 10.1093/nar/21.3.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elias P., Lehman I. R. Interaction of origin binding protein with an origin of replication of herpes simplex virus 1. Proc Natl Acad Sci U S A. 1988 May;85(9):2959–2963. doi: 10.1073/pnas.85.9.2959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FRAENKEL-CONRAT H., SINGER B., TSUGITA A. Purification of viral RNA by means of bentonite. Virology. 1961 May;14:54–58. doi: 10.1016/0042-6822(61)90131-3. [DOI] [PubMed] [Google Scholar]
- Heus H. A., Pardi A. Structural features that give rise to the unusual stability of RNA hairpins containing GNRA loops. Science. 1991 Jul 12;253(5016):191–194. doi: 10.1126/science.1712983. [DOI] [PubMed] [Google Scholar]
- Heus H. A., Uhlenbeck O. C., Pardi A. Sequence-dependent structural variations of hammerhead RNA enzymes. Nucleic Acids Res. 1990 Mar 11;18(5):1103–1108. doi: 10.1093/nar/18.5.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirao I., Ishida M., Watanabe K., Miura K. Unique hairpin structures occurring at the replication origin of phage G4 DNA. Biochim Biophys Acta. 1990 Oct 23;1087(2):199–204. doi: 10.1016/0167-4781(90)90205-g. [DOI] [PubMed] [Google Scholar]
- Hirao I., Nishimura Y., Naraoka T., Watanabe K., Arata Y., Miura K. Extraordinary stable structure of short single-stranded DNA fragments containing a specific base sequence: d(GCGAAAGC). Nucleic Acids Res. 1989 Mar 25;17(6):2223–2231. doi: 10.1093/nar/17.6.2223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirao I., Nishimura Y., Tagawa Y., Watanabe K., Miura K. Extraordinarily stable mini-hairpins: electrophoretical and thermal properties of the various sequence variants of d(GCGAAAGC) and their effect on DNA sequencing. Nucleic Acids Res. 1992 Aug 11;20(15):3891–3896. doi: 10.1093/nar/20.15.3891. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirao I., Yoshizawa S., Miura K. Stabilization of mRNA in an Escherichia coli cell-free translation system. FEBS Lett. 1993 Apr 26;321(2-3):169–172. doi: 10.1016/0014-5793(93)80101-y. [DOI] [PubMed] [Google Scholar]
- James J. K., Tinoco I., Jr The solution structure of a d[C(TTCG)G] DNA hairpin and comparison to the unusually stable RNA analogue. Nucleic Acids Res. 1993 Jul 11;21(14):3287–3293. doi: 10.1093/nar/21.14.3287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katahira M., Sato H., Mishima K., Uesugi S., Fujii S. NMR studies of G:A mismatches in oligodeoxyribonucleotide duplexes modelled after ribozymes. Nucleic Acids Res. 1993 Nov 25;21(23):5418–5424. doi: 10.1093/nar/21.23.5418. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Khan I. M., Coulson J. M. A novel method to stabilise antisense oligonucleotides against exonuclease degradation. Nucleic Acids Res. 1993 Jun 25;21(12):2957–2958. doi: 10.1093/nar/21.12.2957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li Y., Zon G., Wilson W. D. NMR and molecular modeling evidence for a G.A mismatch base pair in a purine-rich DNA duplex. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):26–30. doi: 10.1073/pnas.88.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Michel F., Westhof E. Modelling of the three-dimensional architecture of group I catalytic introns based on comparative sequence analysis. J Mol Biol. 1990 Dec 5;216(3):585–610. doi: 10.1016/0022-2836(90)90386-Z. [DOI] [PubMed] [Google Scholar]
- Ohta T., Sarkar S., Thach R. E. The role of guanosine 5'-triphosphate in the initiation of peptide synthesis. 3. Binding of formylmethionyl-tRNA to ribosomes. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1638–1644. doi: 10.1073/pnas.58.4.1638. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakata T., Hiroaki H., Oda Y., Tanaka T., Ikehara M., Uesugi S. Studies on the structure and stabilizing factor of the CUUCGG hairpin RNA using chemically synthesized oligonucleotides. Nucleic Acids Res. 1990 Jul 11;18(13):3831–3839. doi: 10.1093/nar/18.13.3831. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sklenár V., Miyashiro H., Zon G., Miles H. T., Bax A. Assignment of the 31P and 1H resonances in oligonucleotides by two-dimensional NMR spectroscopy. FEBS Lett. 1986 Nov 10;208(1):94–98. doi: 10.1016/0014-5793(86)81539-3. [DOI] [PubMed] [Google Scholar]
- Summers M. F., Byrd R. A., Gallo K. A., Samson C. J., Zon G., Egan W. Nuclear magnetic resonance and circular dichroism studies of a duplex--single-stranded hairpin loop equilibrium for the oligodeoxyribonucleotide sequence d(CGCGATTCGCG). Nucleic Acids Res. 1985 Sep 11;13(17):6375–6386. doi: 10.1093/nar/13.17.6375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tang J. Y., Temsamani J., Agrawal S. Self-stabilized antisense oligodeoxynucleotide phosphorothioates: properties and anti-HIV activity. Nucleic Acids Res. 1993 Jun 11;21(11):2729–2735. doi: 10.1093/nar/21.11.2729. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsuboi M., Takahashi S., Kyogoku Y., Hayatsu H., Ukita T., Kainosho M. Phosphorus-proton spin-spin coupling and conformation of a dinucleoside phosphate. Science. 1969 Dec 19;166(3912):1504–1505. doi: 10.1126/science.166.3912.1504. [DOI] [PubMed] [Google Scholar]
- Tuerk C., Gauss P., Thermes C., Groebe D. R., Gayle M., Guild N., Stormo G., d'Aubenton-Carafa Y., Uhlenbeck O. C., Tinoco I., Jr CUUCGG hairpins: extraordinarily stable RNA secondary structures associated with various biochemical processes. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1364–1368. doi: 10.1073/pnas.85.5.1364. [DOI] [PMC free article] [PubMed] [Google Scholar]