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. 1978 Jun;75(6):2626–2630. doi: 10.1073/pnas.75.6.2626

Isolation and characterization of a highly repetitious inverted terminal repeat sequence from Oxytricha macronuclear DNA.

G Herrick, R D Wesley
PMCID: PMC392615  PMID: 96439

Abstract

The low-complexity "gene-sized" linear DNA duplexes of the Oxytricha macronucleus sport short inverted terminal repeats; thus, each single strand is capable of forming a circle held together by a duplex "neck" [Wesley, R. D. (1975) Porc. Natl. Acad. Sci. USA 72, 678--682]. We have isolated necks from total, circularized, single-stranded macronuclear DNA by treatment with nuclease S1. Necks represent at least 2.2% of the total DNA, are homogeneous in size (23 base pairs), melt at 55 degrees in 0.18 M Na+, and reassociate extremely rapidly at 22 degrees (Cot1/2 = 1.1 X 10(-5) mol-liter-1.sec) to form hybrid necks of the same thermal stability. From these and other results, we conclude that all necks on all the many thousands of different single-stranded circles are the same. The neck sequence is therefore highly repetitious--found in multiple copies (as inverted terminal repeats at flush duplex ends and probably also internally) on each natural "gene-sized" macronuclear DNA molecule--implying the possible participation of this sequence both in the general vegetative metabolism of macronuclear DNA and in the pre-vegetative process whereby macronuclear DNA is excised from the total Oxytricha genome.

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Selected References

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

  1. Englund P. T. Analysis of nucleotide sequences at 3' termini of duplex deoxyribonucleic acid with the use of the T4 deoxyribonucleic acid polymerase. J Biol Chem. 1971 May 25;246(10):3269–3276. [PubMed] [Google Scholar]
  2. Hayward G. S., Jacob R. J., Wadsworth S. C., Roizman B. Anatomy of herpes simplex virus DNA: evidence for four populations of molecules that differ in the relative orientations of their long and short components. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4243–4247. doi: 10.1073/pnas.72.11.4243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hsu M. T., Davidson N. Electron microscope heteroduplex study of the heterogeneity of Mu phage and prophage DNA. Virology. 1974 Mar;58(1):229–239. doi: 10.1016/0042-6822(74)90157-3. [DOI] [PubMed] [Google Scholar]
  4. Kleckner N. Translocatable elements in procaryotes. Cell. 1977 May;11(1):11–23. doi: 10.1016/0092-8674(77)90313-0. [DOI] [PubMed] [Google Scholar]
  5. Lauth M. R., Spear B. B., Heumann J., Prescott D. M. DNA of ciliated protozoa: DNA sequence diminution during macronuclear development of Oxytricha. Cell. 1976 Jan;7(1):67–74. doi: 10.1016/0092-8674(76)90256-7. [DOI] [PubMed] [Google Scholar]
  6. Lawn R. M. Gene-sized DNA molecules of the Oxytricha macronucleus have the same terminal sequence. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4325–4328. doi: 10.1073/pnas.74.10.4325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  8. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Michelson A. M., Massoulié J., Guschlbauer W. Synthetic polynucleotides. Prog Nucleic Acid Res Mol Biol. 1967;6:83–141. doi: 10.1016/s0079-6603(08)60525-5. [DOI] [PubMed] [Google Scholar]
  10. Nossal N. G., Hershfield M. S. Nuclease activity in a fragment of bacteriophage T4 deoxyribonucleic acid polymerase induced by the amber mutant am B22. J Biol Chem. 1971 Sep 10;246(17):5414–5426. [PubMed] [Google Scholar]
  11. Pellegrini M., Manning J., Davidson N. Sequence arrangement of the rDNA of Drosophila melanogaster. Cell. 1977 Feb;10(2):213–214. doi: 10.1016/0092-8674(77)90215-x. [DOI] [PubMed] [Google Scholar]
  12. Perlman S., Phillips C., Bishop J. O. A study of foldback DNA. Cell. 1976 May;8(1):33–42. doi: 10.1016/0092-8674(76)90182-3. [DOI] [PubMed] [Google Scholar]
  13. RICHARDSON C. C., LEHMAN I. R., KORNBERG A. A DEOXYRIBONUCLEIC ACID PHOSPHATASE-EXONUCLEASE FROM ESCHERICHIA COLI. II. CHARACTERIZATION OF THE EXONUCLEASE ACTIVITY. J Biol Chem. 1964 Jan;239:251–258. [PubMed] [Google Scholar]
  14. Schmid C. W., Manning J. E., Davidson N. Inverted repeat sequences in the Drosophila genome. Cell. 1975 Jun;5(2):159–172. doi: 10.1016/0092-8674(75)90024-0. [DOI] [PubMed] [Google Scholar]
  15. Shenk T. E., Rhodes C., Rigby P. W., Berg P. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40. Proc Natl Acad Sci U S A. 1975 Mar;72(3):989–993. doi: 10.1073/pnas.72.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sutton W. D. A crude nuclease preparation suitable for use in DNA reassociation experiments. Biochim Biophys Acta. 1971 Jul 29;240(4):522–531. doi: 10.1016/0005-2787(71)90709-x. [DOI] [PubMed] [Google Scholar]
  17. Wesley R. D. Inverted repetitious sequences in the macronuclear DNA of hypotrichous ciliates. Proc Natl Acad Sci U S A. 1975 Feb;72(2):678–682. doi: 10.1073/pnas.72.2.678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wetmur J. G., Davidson N. Kinetics of renaturation of DNA. J Mol Biol. 1968 Feb 14;31(3):349–370. doi: 10.1016/0022-2836(68)90414-2. [DOI] [PubMed] [Google Scholar]
  19. Yansura D. G., Goeddel D. V., Cribbs D. L., Caruthers M. H. Studies of gene control regions. III. Binding of synthetic and modified synthetic lac operator DNAs to lactose repressor. Nucleic Acids Res. 1977 Mar;4(3):723–737. doi: 10.1093/nar/4.3.723. [DOI] [PMC free article] [PubMed] [Google Scholar]

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