Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1979;6(6):2089–2107. doi: 10.1093/nar/6.6.2089

The circular dichroism and X-ray diffraction of DNA condensed from ethanolic solutions.

D M Gray, S P Edmondson, D Lang, M Vaughan
PMCID: PMC327838  PMID: 572544

Abstract

It is known that DNA in aqueous-ethanol solutions undergoes a B to A conformational change between 60% and 80% (w/w) ethanol. We have found that precipitates formed by adding salt to DNA in 60% and 80% ethanolic solutions can be very different. DNA precipitated from 60% ethanol forms a fine condensate that only slowly settles out of suspension and shows a characteristic differential scattering of circularly polarized light at long wavelengths. DNA precipitated from 80% ethanol forms a flocculent aggregate that exhibits the CD spectral features of the A conformation. Data from circular dichroism spectra of natural and synthetic nucleic acids and from X-ray diffraction patterns of the precipitates show that DNA molecules precipitated from 60% and 80% ethanol are, respectively, in the B and A conformation. Therefore, the different secondary conformations of DNA in ethanolic solutions are maintained during precipitation under these conditions. These results are of general importance for the preparation and study of condensed forms of DNA, since a relatively small change in the extent of dehydration can change the secondary conformation of DNA and markedly affect the character of a subsequent precipitate.

Full text

PDF
2089

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnott S., Hukins D. W., Dover S. D., Fuller W., Hodgson A. R. Structures of synthetic polynucleotides in the A-RNA and A'-RNA conformations: x-ray diffraction analyses of the molecular conformations of polyadenylic acid--polyuridylic acid and polyinosinic acid--polycytidylic acid. J Mol Biol. 1973 Dec 5;81(2):107–122. doi: 10.1016/0022-2836(73)90183-6. [DOI] [PubMed] [Google Scholar]
  2. Arnott S., Hukins D. W. Refinement of the structure of B-DNA and implications for the analysis of x-ray diffraction data from fibers of biopolymers. J Mol Biol. 1973 Dec 5;81(2):93–105. doi: 10.1016/0022-2836(73)90182-4. [DOI] [PubMed] [Google Scholar]
  3. Arnott S. The geometry of nucleic acids. Prog Biophys Mol Biol. 1970;21:265–319. doi: 10.1016/0079-6107(70)90027-1. [DOI] [PubMed] [Google Scholar]
  4. Cooper P. J., Hamilton L. D. The A-B conformational change in the sodium salt of DNA. J Mol Biol. 1966 Apr;16(2):562–563. doi: 10.1016/s0022-2836(66)80193-6. [DOI] [PubMed] [Google Scholar]
  5. Damaschun H., Damaschun G., Becker M., Buder E., Misselwitz R., Zirwer D. Study of DNA-spermine interactions by use of small-angle and wide-angle X-ray scattering and circular dichroism. Nucleic Acids Res. 1978 Oct;5(10):3801–3809. doi: 10.1093/nar/5.10.3801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Earnshaw W., Casjens S., Harrison S. C. Assembly of the head of bacteriophage P22: x-ray diffraction from heads, proheads and related structures. J Mol Biol. 1976 Jun 25;104(2):387–410. doi: 10.1016/0022-2836(76)90278-3. [DOI] [PubMed] [Google Scholar]
  7. Eickbush T. H., Moudrianakis E. N. The compaction of DNA helices into either continuous supercoils or folded-fiber rods and toroids. Cell. 1978 Feb;13(2):295–306. doi: 10.1016/0092-8674(78)90198-8. [DOI] [PubMed] [Google Scholar]
  8. Evdokimov Y. M., Pyatigorskaya T. L., Kadikov V. A., Polyvtsev O. F., Doskocil J., Koudelka J., Varshavsky Y. M. A compact form of double-stranded RNA in solutions containing poly(ethyleneglycol). Nucleic Acids Res. 1976 Jun;3(6):1533–1547. doi: 10.1093/nar/3.6.1533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. FULLER W., WILKINS M. H., WILSON H. R., HAMILTON L. D. THE MOLECULAR CONFIGURATION OF DEOXYRIBONUCLEIC ACID. IV. X-RAY DIFFRACTION STUDY OF THE A FORM. J Mol Biol. 1965 May;12:60–76. doi: 10.1016/s0022-2836(65)80282-0. [DOI] [PubMed] [Google Scholar]
  10. Giannoni G., Padden F. J., Jr, Keith H. D. Crystallization of DNA from dilute solution. Proc Natl Acad Sci U S A. 1969 Mar;62(3):964–971. doi: 10.1073/pnas.62.3.964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Girod J. C., Johnson W. C., Jr, Huntington S. K., Maestre M. F. Conformation of deoxyribonucleic acid in alcohol solutions. Biochemistry. 1973 Dec 4;12(25):5092–5096. doi: 10.1021/bi00749a011. [DOI] [PubMed] [Google Scholar]
  12. Goodwin D. C., Brahms J. Form of DNA and the nature of interactions with proteins in chromatin. Nucleic Acids Res. 1978 Mar;5(3):835–850. doi: 10.1093/nar/5.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gosule L. C., Schellman J. A. DNA condensation with polyamines I. Spectroscopic studies. J Mol Biol. 1978 May 25;121(3):311–326. doi: 10.1016/0022-2836(78)90366-2. [DOI] [PubMed] [Google Scholar]
  14. Gray D. M., Ratliff R. L. Circular dichroism spectra of poly[d(AC):d(GT)], poly[r(AC):r(GU)], and hybrids poly[d(AC):r(GU)] and poly[r(AC):d(GT)] in the presence of ethanol. Biopolymers. 1975 Mar;14(3):487–498. doi: 10.1002/bip.1975.360140305. [DOI] [PubMed] [Google Scholar]
  15. Gray D. M., Ratliff R. L., Williams D. L. The circular dichroism of poly d(AAT):d(ATT) and poly r(AAU):r(AUU). A test of the first-neighbor hypothesis. Biopolymers. 1973 Jun;12(6):1233–1245. doi: 10.1002/bip.1973.360120604. [DOI] [PubMed] [Google Scholar]
  16. Gray D. M., Taylor T. N., Lang D. Dehydrated circular DNA: circular dichroism of molecules in ethanolic solutions. Biopolymers. 1978 Jan;17(1):145–157. doi: 10.1002/bip.1978.360170111. [DOI] [PubMed] [Google Scholar]
  17. Hirschman S. Z., Felsenfeld G. Determination of DNA composition and concentration by spectral analysis. J Mol Biol. 1966 Apr;16(2):347–358. doi: 10.1016/s0022-2836(66)80178-x. [DOI] [PubMed] [Google Scholar]
  18. Ivanov V. I., Minchenkova L. E., Minyat E. E., Frank-Kamenetskii M. D., Schyolkina A. K. The B to A transition of DNA in solution. J Mol Biol. 1974 Aug 25;87(4):817–833. doi: 10.1016/0022-2836(74)90086-2. [DOI] [PubMed] [Google Scholar]
  19. Jordan C. F., Lerman L. S., Venable J. H. Structure and circular dichroism of DNA in concentrated polymer solutions. Nat New Biol. 1972 Mar 22;236(64):67–70. doi: 10.1038/newbio236067a0. [DOI] [PubMed] [Google Scholar]
  20. Lang D. Regular superstructures of purified DNA in ethanolic solutions. J Mol Biol. 1973 Aug 5;78(2):247–254. doi: 10.1016/0022-2836(73)90113-7. [DOI] [PubMed] [Google Scholar]
  21. Lang D., Taylor T. N., Dobyan D. C., Gray D. M. Dehydrated circular DNA: electron microscopy of ethanol-condensed molecules. J Mol Biol. 1976 Sep 5;106(1):97–107. doi: 10.1016/0022-2836(76)90302-8. [DOI] [PubMed] [Google Scholar]
  22. Lerman L. S., Wilkerson L. S., Venable J. H., Jr, Robinson B. H. DNA packing in single crystals inferred from freeze-fracture-etch replicas. J Mol Biol. 1976 Dec;108(2):271–293. doi: 10.1016/s0022-2836(76)80121-0. [DOI] [PubMed] [Google Scholar]
  23. Levitt M. How many base-pairs per turn does DNA have in solution and in chromatin? Some theoretical calculations. Proc Natl Acad Sci U S A. 1978 Feb;75(2):640–644. doi: 10.1073/pnas.75.2.640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MARVIN D. A., SPENCER M., WILKINS M. H., HAMILTON L. D. The molecular configuration of deoxyribonucleic acid. III. X-ray diffraction study of the C form of the lithium salt. J Mol Biol. 1961 Oct;3:547–565. doi: 10.1016/s0022-2836(61)80021-1. [DOI] [PubMed] [Google Scholar]
  25. Maniatis T., Venable J. H., Jr, Lerman L. S. The structure of psi DNA. J Mol Biol. 1974 Mar 25;84(1):37–64. doi: 10.1016/0022-2836(74)90211-3. [DOI] [PubMed] [Google Scholar]
  26. RUBENSTEIN I., THOMAS C. A., Jr, HERSHEY A. D. The molecular weights of T2 bacteriophage DNA and its first and second breakage products. Proc Natl Acad Sci U S A. 1961 Aug;47:1113–1122. doi: 10.1073/pnas.47.8.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shin Y. A., Eichhorn G. L. Reversible change in psi structure of DNA-poly(Lys) complexes induced by metal binding. Biopolymers. 1977 Jan;16(1):225–230. doi: 10.1002/bip.1977.360160117. [DOI] [PubMed] [Google Scholar]
  28. Wells R. D., Larson J. E., Grant R. C., Shortle B. E., Cantor C. R. Physicochemical studies on polydeoxyribonucleotides containing defined repeating nucleotide sequences. J Mol Biol. 1970 Dec 28;54(3):465–497. doi: 10.1016/0022-2836(70)90121-x. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES