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. 1985 Jul;4(7):1881–1886. doi: 10.1002/j.1460-2075.1985.tb03864.x

Hairpin plasmid--a novel linear DNA of perfect hairpin structure.

Y Kikuchi, K Hirai, N Gunge, F Hishinuma
PMCID: PMC554431  PMID: 2992950

Abstract

The terminal structures of deletion derivatives of linear DNA killer plasmid from yeast were analyzed. The yeast Kluyveromyces lactis harbors two unique double-stranded linear DNA killer plasmids, pGKL1 of 8.9 kb and pGKL2 of 13.4 kb. The killer toxin and the resistance to the killer are coded by pGKL1, while pGKL2 is required for the maintenance of pGKL1 in the cell. When the pGKL plasmids from K. lactis were transferred into Saccharomyces cerevisiae by transformation, non-killer transformants harboring pGKL2 and new plasmids, F1 of 7.8 kb and F2 of 3.9 kb, were obtained. F2 was shown to be a linear DNA arising from a 5-kb deletion of the right part of pGKL1. F1 was an inverted dimer of F2. Here we show that F2 has two different terminal structures: one end has a protein attached at the 5' terminus whereas the two strands of duplex are linked together at the other end, thus forming a hairpin structure. This is a novel type of autonomously replicating DNA molecule.

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Selected References

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

  1. Arrand J. R., Roberts R. J. The nucleotide sequences at the termini of adenovirus-2 DNA. J Mol Biol. 1979 Mar 15;128(4):577–594. doi: 10.1016/0022-2836(79)90294-8. [DOI] [PubMed] [Google Scholar]
  2. Blackburn E. H. The molecular structure of centromeres and telomeres. Annu Rev Biochem. 1984;53:163–194. doi: 10.1146/annurev.bi.53.070184.001115. [DOI] [PubMed] [Google Scholar]
  3. Carusi E. A. Evidence for blocked 5'-termini in human adenovirus DNA. Virology. 1977 Jan;76(1):380–394. doi: 10.1016/0042-6822(77)90310-5. [DOI] [PubMed] [Google Scholar]
  4. Challberg M. D., Kelly T. J. Eukaryotic DNA replication: viral and plasmid model systems. Annu Rev Biochem. 1982;51:901–934. doi: 10.1146/annurev.bi.51.070182.004345. [DOI] [PubMed] [Google Scholar]
  5. Desiderio S. V., Kelly T. J., Jr Structure of the linkage between adenovirus DNA and the 55,000 molecular weight terminal protein. J Mol Biol. 1981 Jan 15;145(2):319–337. doi: 10.1016/0022-2836(81)90208-4. [DOI] [PubMed] [Google Scholar]
  6. Escarmís C., Salas M. Nucleotide sequence at the termini of the DNA of Bacillus subtilis phage phi 29. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1446–1450. doi: 10.1073/pnas.78.3.1446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gunge N., Murata K., Sakaguchi K. Transformation of Saccharomyces cerevisiae with linear DNA killer plasmids from Kluyveromyces lactis. J Bacteriol. 1982 Jul;151(1):462–464. doi: 10.1128/jb.151.1.462-464.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gunge N., Sakaguchi K. Intergeneric transfer of deoxyribonucleic acid killer plasmids, pGKl1 and pGKl2, from Kluyveromyces lactis into Saccharomyces cerevisiae by cell fusion. J Bacteriol. 1981 Jul;147(1):155–160. doi: 10.1128/jb.147.1.155-160.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gunge N., Tamaru A., Ozawa F., Sakaguchi K. Isolation and characterization of linear deoxyribonucleic acid plasmids from Kluyveromyces lactis and the plasmid-associated killer character. J Bacteriol. 1981 Jan;145(1):382–390. doi: 10.1128/jb.145.1.382-390.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunge N. Yeast DNA plasmids. Annu Rev Microbiol. 1983;37:253–276. doi: 10.1146/annurev.mi.37.100183.001345. [DOI] [PubMed] [Google Scholar]
  11. Harding N. E., Ito J. DNA replication of bacteriophage phi 29: characterization of the intermediates and location of the termini of replication. Virology. 1980 Jul 30;104(2):323–338. doi: 10.1016/0042-6822(80)90337-2. [DOI] [PubMed] [Google Scholar]
  12. Harding N. E., Ito J., David G. S. Identification of the protein firmly bound to the ends of bacteriophage phi 29 DNA. Virology. 1978 Feb;84(2):279–292. doi: 10.1016/0042-6822(78)90248-9. [DOI] [PubMed] [Google Scholar]
  13. Hirochika H., Nakamura K., Sakaguchi K. A linear DNA plasmid from Streptomyces rochei with an inverted terminal repetition of 614 base pairs. EMBO J. 1984 Apr;3(4):761–766. doi: 10.1002/j.1460-2075.1984.tb01881.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hirochika H., Sakaguchi K. Analysis of linear plasmids isolated from Streptomyces: association of protein with the ends of the plasmid DNA. Plasmid. 1982 Jan;7(1):59–65. doi: 10.1016/0147-619x(82)90027-0. [DOI] [PubMed] [Google Scholar]
  15. Hishinuma F., Nakamura K., Hirai K., Nishizawa R., Gunge N., Maeda T. Cloning and nucleotide sequences of the linear DNA killer plasmids from yeast. Nucleic Acids Res. 1984 Oct 11;12(19):7581–7597. doi: 10.1093/nar/12.19.7581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Inciarte M. R., Salas M., Sogo J. M. Structure of replicating DNA molecules of Bacillus subtilis bacteriophage phi 29. J Virol. 1980 Apr;34(1):187–199. doi: 10.1128/jvi.34.1.187-199.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito J. Bacteriophage phi29 terminal protein: its association with the 5' termini of the phi29 genome. J Virol. 1978 Dec;28(3):895–904. doi: 10.1128/jvi.28.3.895-904.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kemble R. J., Thompson R. D. S1 and S2, the linear mitochondrial DNAs present in a male sterile line of maize, possess terminally attached proteins. Nucleic Acids Res. 1982 Dec 20;10(24):8181–8190. doi: 10.1093/nar/10.24.8181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kikuchi Y., Hirai K., Hishinuma F. The yeast linear DNA killer plasmids, pGKL1 and pGKL2, possess terminally attached proteins. Nucleic Acids Res. 1984 Jul 25;12(14):5685–5692. doi: 10.1093/nar/12.14.5685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lichy J. H., Nagata K., Friefeld B. R., Enomoto T., Field J., Guggenheimer R. A., Ikeda J. E., Horwitz M. S., Hurwitz J. Isolation of proteins involved in the replication of adenoviral DNA in vitro. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):731–740. doi: 10.1101/sqb.1983.047.01.084. [DOI] [PubMed] [Google Scholar]
  21. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  22. Niwa O., Sakaguchi K., Gunge N. Curing of the killer deoxyribonucleic acid plasmids of Kluyveromyces lactis. J Bacteriol. 1981 Dec;148(3):988–990. doi: 10.1128/jb.148.3.988-990.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Prieto I., Lázaro J. M., García J. A., Hermoso J. M., Salas M. Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1639–1643. doi: 10.1073/pnas.81.6.1639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pring D. R., Levings C. S., Hu W. W., Timothy D. H. Unique DNA associated with mitochondria in the "S"-type cytoplasm of male-sterile maize. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2904–2908. doi: 10.1073/pnas.74.7.2904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rekosh D. M., Russell W. C., Bellet A. J., Robinson A. J. Identification of a protein linked to the ends of adenovirus DNA. Cell. 1977 Jun;11(2):283–295. doi: 10.1016/0092-8674(77)90045-9. [DOI] [PubMed] [Google Scholar]
  26. Salas M., Mellado R. P., Viñuela E. Characterization of a protein covalently linked to the 5' termini of the DNA of Bacillus subtilis phage phi29. J Mol Biol. 1978 Feb 25;119(2):269–291. doi: 10.1016/0022-2836(78)90438-2. [DOI] [PubMed] [Google Scholar]
  27. Shinagawa M., Padmanabhan R. Nucleotide sequence at the inverted terminal repetition of adenovirus type 2 DNA. Biochem Biophys Res Commun. 1979 Apr 13;87(3):671–678. doi: 10.1016/0006-291x(79)92011-4. [DOI] [PubMed] [Google Scholar]
  28. Silberklang M., Gillum A. M., RajBhandary U. L. Use of in vitro 32P labeling in the sequence analysis of nonradioactive tRNAs. Methods Enzymol. 1979;59:58–109. doi: 10.1016/0076-6879(79)59072-7. [DOI] [PubMed] [Google Scholar]
  29. Sogo J. M., García J. A., Peñalva M. A., Salas M. Structure of protein-containing replicative intermediates of Bacillus subtilis phage phi 29 DNA. Virology. 1982 Jan 15;116(1):1–18. doi: 10.1016/0042-6822(82)90398-1. [DOI] [PubMed] [Google Scholar]
  30. Sor F., Wésolowski M., Fukuhara H. Inverted terminal repetitions of the two linear DNA associated with the killer character of the yeast Kluyveromyces lactis. Nucleic Acids Res. 1983 Aug 11;11(15):5037–5044. doi: 10.1093/nar/11.15.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stark M. J., Mileham A. J., Romanos M. A., Boyd A. Nucleotide sequence and transcription analysis of a linear DNA plasmid associated with the killer character of the yeast Kluyveromyces lactis. Nucleic Acids Res. 1984 Aug 10;12(15):6011–6030. doi: 10.1093/nar/12.15.6011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Steenbergh P. H., Maat J., van Ormondt H., Sussenbach J. S. The nucleotide sequence at the termini of adenovirus type 5 DNA. Nucleic Acids Res. 1977 Dec;4(12):4371–4389. doi: 10.1093/nar/4.12.4371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stillman B. W. The replication of adenovirus DNA with purified proteins. Cell. 1983 Nov;35(1):7–9. doi: 10.1016/0092-8674(83)90201-5. [DOI] [PubMed] [Google Scholar]
  34. Sugisaki Y., Gunge N., Sakaguchi K., Yamasaki M., Tamura G. Characterization of a novel killer toxin encoded by a double-stranded linear DNA plasmid of Kluyveromyces lactis. Eur J Biochem. 1984 Jun 1;141(2):241–245. doi: 10.1111/j.1432-1033.1984.tb08183.x. [DOI] [PubMed] [Google Scholar]
  35. Sugisaki Y., Gunge N., Sakaguchi K., Yamasaki M., Tamura G. Kluyveromyces lactis killer toxin inhibits adenylate cyclase of sensitive yeast cells. Nature. 1983 Aug 4;304(5925):464–466. doi: 10.1038/304464a0. [DOI] [PubMed] [Google Scholar]
  36. Yehle C. O. Genome-linked protein associated with the 5' termini of bacteriophage phi29 DNA. J Virol. 1978 Sep;27(3):776–783. doi: 10.1128/jvi.27.3.776-783.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yoshikawa H., Friedmann T., Ito J. Nucleotide sequences at the termini of phi 29 DNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1336–1340. doi: 10.1073/pnas.78.3.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]

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