Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1975 Mar;2(3):431–446. doi: 10.1093/nar/2.3.431

Identification and separation of components of calf thymus DNA using a CsCl-netropsin density gradient

H Votavová 1, J Šponar 1
PMCID: PMC342848  PMID: 805418

Abstract

Calf thymus DNA containing satellite components of various densities was used as a model to study the effect of netropsin on the density of DNA in a CsCl gradient. The binding of netropsin resulted in a decrease in density which depended upon the quantity of netropsin added and on the average composition of the DNA. Differences in density of DNA components were higher in CsCl - netropsin gradients than in simple CsCl gradients. By use of netropsin a main band and four satellite bands could be differentiated in calf thymus DNA. Satellite DNA's were isolated using preparative CsCl - netropsin gradient centrifugation and were characterised by density and homogeneity in native and in reassociated state. Two of the satellite components, with densities of 1.722 and 1.714 g/cm3, are probably of homogenous sequence, the other two components of densities 1.709 and 1.705 g/cm3 appear to be heterogeneous.

Full text

PDF
431

Selected References

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

  1. Blumenfeld M., Fox A. S., Forrest H. S. A family of three related satellite DNAs in Drosophila virilis. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2772–2775. doi: 10.1073/pnas.70.10.2772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botchan M. R. Bovine satellite I DNA consists of repetitive units 1,400 base pairs in length. Nature. 1974 Sep 27;251(5473):288–292. doi: 10.1038/251288a0. [DOI] [PubMed] [Google Scholar]
  3. Corneo G., Ginelli E., Polli E. Different satellite deoxyribonucleic acids of guinea pig and ox. Biochemistry. 1970 Mar 31;9(7):1565–1571. doi: 10.1021/bi00809a014. [DOI] [PubMed] [Google Scholar]
  4. DAVIDSON N., WIDHOLM J., NANDI U. S., JENSEN R., OLIVERA B. M., WANG J. C. PREPARATION AND PROPERTIES OF NATIVE CRAB DAT. Proc Natl Acad Sci U S A. 1965 Jan;53:111–118. doi: 10.1073/pnas.53.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Filipski J., Thiery J. P., Bernardi G. An analysis of the bovine genome by Cs2SO4-Ag density gradient centrifugation. J Mol Biol. 1973 Oct 15;80(1):177–197. doi: 10.1016/0022-2836(73)90240-4. [DOI] [PubMed] [Google Scholar]
  6. Kersten W., Kersten H., Szybalski W. Physicochemical properties of complexes between deoxyribonucleic acid and antibiotics which affect ribonucleic acid synthesis (actinomycin, daunomycin, cinerubin, nogalamycin, chormomycin, mithramycin, and olivomycin). Biochemistry. 1966 Jan;5(1):236–244. doi: 10.1021/bi00865a031. [DOI] [PubMed] [Google Scholar]
  7. Kurnit D. M., Shafit B. R., Maio J. J. Multiple satellite deoxyribonucleic acids in the calf and their relation to the sex chromosomes. J Mol Biol. 1973 Dec 15;81(3):273–284. doi: 10.1016/0022-2836(73)90141-1. [DOI] [PubMed] [Google Scholar]
  8. Luck G., Triebel H., Waring M., Zimmer C. Conformation dependent binding of netropsin and distamycin to DNA and DNA model polymers. Nucleic Acids Res. 1974 Mar;1(3):503–530. doi: 10.1093/nar/1.3.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. MANDELL J. D., HERSHEY A. D. A fractionating column for analysis of nucleic acids. Anal Biochem. 1960 Jun;1:66–77. doi: 10.1016/0003-2697(60)90020-8. [DOI] [PubMed] [Google Scholar]
  10. Peacock W. J., Brutlag D., Goldring E., Appels R., Hinton C. W., Lindsley D. L. The organization of highly repeated DNA sequences in Drosophila melanogaster chromosomes. Cold Spring Harb Symp Quant Biol. 1974;38:405–416. doi: 10.1101/sqb.1974.038.01.043. [DOI] [PubMed] [Google Scholar]
  11. Polli E., Ginelli E., Bianchi P., Corneo G. Renaturation of calf thymus satellite DNA. J Mol Biol. 1966 May;17(1):305–308. doi: 10.1016/s0022-2836(66)80114-6. [DOI] [PubMed] [Google Scholar]
  12. Reinert K. E. Adenosine-thymidine cluster-specific elongation and stiffening of DNA induced by the oligopeptide antibiotic netropsin. J Mol Biol. 1972 Dec 30;72(3):593–607. doi: 10.1016/0022-2836(72)90178-7. [DOI] [PubMed] [Google Scholar]
  13. SCHILDKRAUT C. L., MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J Mol Biol. 1962 Jun;4:430–443. doi: 10.1016/s0022-2836(62)80100-4. [DOI] [PubMed] [Google Scholar]
  14. VINOGRAD J., HEARST J. E. Equilibrium sedimentation of macromolecules and viruses in a density gradient. Fortschr Chem Org Naturst. 1962;20:373–422. [PubMed] [Google Scholar]
  15. Votavová H., Sponar J. Reassociation kinetics of three satellite components of calf thymus DNA. Nucleic Acids Res. 1975 Feb;2(2):185–196. doi: 10.1093/nar/2.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Votavová H., Sponar J., Sormová Z. Isolation and properties of rapidly renaturing fractions of DNA from calf tissues. Eur J Biochem. 1970 Feb;12(2):208–216. doi: 10.1111/j.1432-1033.1970.tb00839.x. [DOI] [PubMed] [Google Scholar]
  17. Wartell R. M., Larson J. E., Wells R. D. Netropsin. A specific probe for A-T regions of duplex deoxyribonucleic acid. J Biol Chem. 1974 Nov 10;249(21):6719–6731. [PubMed] [Google Scholar]
  18. Yasmineh W. G., Yunis J. J. Satellite DNA in calf heterochromatin. Exp Cell Res. 1971 Jan;64(1):41–48. doi: 10.1016/0014-4827(71)90190-x. [DOI] [PubMed] [Google Scholar]
  19. Zimmer C., Luck G. Stability and dissociation of the DNA complexes with distamycin A and Netropsin in the presence of organic solvents, urea and high salt concentration. Biochim Biophys Acta. 1972 Dec 22;287(3):376–385. doi: 10.1016/0005-2787(72)90281-x. [DOI] [PubMed] [Google Scholar]
  20. Zimmer C., Puschendorf B., Grunicke H., Chandra P., Venner H. Influence of netropsin and distamycin A on the secondary structure and template activity of DNA. Eur J Biochem. 1971 Jul 29;21(2):269–278. doi: 10.1111/j.1432-1033.1971.tb01466.x. [DOI] [PubMed] [Google Scholar]
  21. Zimmer C., Reinert K. E., Luck G., Wähnert U., Löber G., Thrum H. Interaction of the oligopeptide antibiotics netropsin and distamycin A with nucleic acids. J Mol Biol. 1971 May 28;58(1):329–348. doi: 10.1016/0022-2836(71)90250-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES