Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1985 Jan;53(1):25–31. doi: 10.1128/jvi.53.1.25-31.1985

Transfection of REP- mycoplasmas with viral single-stranded DNA.

T L Sladek, J Maniloff
PMCID: PMC254973  PMID: 3965744

Abstract

Double-stranded DNA from mycoplasma virus L2 can transfect Acholeplasma laidlawii cells in the presence of polyethylene glycol (T. L. Sladek and J. Maniloff, J. Bacteriol. 155:734-741, 1983). We report here that both single-stranded DNA and double-stranded replicative form DNA, from the single-stranded DNA mycoplasma virus L51, are also infectious in this system. For both DNAs transfection frequencies were in the range of 10(-8) transfectants per DNA molecule and 10(-3) transfectants per CFU. An unexpected finding was that both DNAs could transfect A. laidlawii strain REP-, a variant which is a nonpermissive host for single-stranded DNA mycoplasma viruses due to a block in viral DNA replication (Nowak et al., J. Bacteriol. 127:832-836, 1976). The number of viruses produced by transfected REP- cells was comparable to the number produced by both transfected and infected wild-type cells. Therefore, transfected L51 DNAs are able to bypass the replication block in REP- cells that occurs when these cells are infected by L51 virions.

Full text

PDF
25

Images in this article

Selected References

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

  1. Beard P., Morrow J. F., Berg P. Cleavage of circular, superhelical simian virus 40 DNA to a linear duplex by S1 nuclease. J Virol. 1973 Dec;12(6):1303–1313. doi: 10.1128/jvi.12.6.1303-1313.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cerone-McLernon A. M., Furness G. The preparation of transforming DNA from Mycoplasma hominis strain Sprott tetr and quantitative studies of the factors affecting the genetic transformation of Mycoplasma salivarium strain S9 tets to tetracycline resistance. Can J Microbiol. 1980 Sep;26(9):1147–1152. doi: 10.1139/m80-189. [DOI] [PubMed] [Google Scholar]
  3. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Das J., Maniloff J. Replication of mycoplasmavirus MVL51: I. Replicative intermediates. Biochem Biophys Res Commun. 1975 Sep 16;66(2):599–605. doi: 10.1016/0006-291x(75)90552-5. [DOI] [PubMed] [Google Scholar]
  5. Das J., Maniloff J. Replication of mycoplasmavirus MVL51: attachment of MVL51 parental DNA to host cell membrane. Proc Natl Acad Sci U S A. 1976 May;73(5):1489–1493. doi: 10.1073/pnas.73.5.1489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Folsome C. E. Deoxyribonucleate binding and transformation in Mycoplasma laidlawii. J Gen Microbiol. 1968 Jan;50(1):43–53. doi: 10.1099/00221287-50-1-43. [DOI] [PubMed] [Google Scholar]
  7. Fraser D., Fleischmann C. Interaction of mycoplasma with viruses. I. Primary adsorption of virus is ionic in mechanism. J Virol. 1974 May;13(5):1067–1074. doi: 10.1128/jvi.13.5.1067-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Germond J. E., Vogt V. M., Hirt B. Characterization of the single-strand-specific nuclease S1 activity on double-stranded supercoiled polyoma DNA. Eur J Biochem. 1974 Apr 16;43(3):591–600. doi: 10.1111/j.1432-1033.1974.tb03446.x. [DOI] [PubMed] [Google Scholar]
  9. Gourlay R. N. Mycoplasmatales virus-laidlawii 2, a new virus isolated from Acholeplasma laidlawii. J Gen Virol. 1971 Jul;12(1):65–67. doi: 10.1099/0022-1317-12-1-65. [DOI] [PubMed] [Google Scholar]
  10. Gourlay R. N., Wyld S. G. Isolation of mycoplasmatales virus-laidlawii 3, a new virus infecting Acholeplasma laidlawii. J Gen Virol. 1973 May;19(2):279–283. doi: 10.1099/0022-1317-19-2-279. [DOI] [PubMed] [Google Scholar]
  11. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Haberer K., Klotz G., Maniloff J., Kleinschmidt A. K. Structural and biological properties of mycoplasmavirus MVL3: an unusual virus-procaryote interaction. J Virol. 1979 Oct;32(1):268–275. doi: 10.1128/jvi.32.1.268-275.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Haberer K., Maniloff J. Adsorption of the tailed mycoplasma virus L3 to cell membranes. J Virol. 1982 Feb;41(2):501–507. doi: 10.1128/jvi.41.2.501-507.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Haberer K., Maniloff J. Virus and host cell DNA syntheses during infection of Acholeplasma laidlawii by MVL3, a nonlytic cytocidal mycoplasmavirus. J Virol. 1980 Feb;33(2):671–679. doi: 10.1128/jvi.33.2.671-679.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kornberg A., Scott J. F., Bertsch L. L. ATP utilization by rep protein in the catalytic separation of DNA strands at a replicating fork. J Biol Chem. 1978 May 10;253(9):3298–3304. [PubMed] [Google Scholar]
  16. Liss A., Maniloff J. Infection of Acholeplasma laidlawii by MVL51 virus. Virology. 1973 Sep;55(1):118–126. doi: 10.1016/s0042-6822(73)81013-x. [DOI] [PubMed] [Google Scholar]
  17. Liss A., Maniloff J. Isolation of Mycoplasmatales viruses and characterization of MVL1, MVL52, and MVG51. Science. 1971 Aug 20;173(3998):725–727. doi: 10.1126/science.173.3998.725. [DOI] [PubMed] [Google Scholar]
  18. Maniloff J., Haberer K., Gourlay R. N., Das J., Cole R. Mycoplasma viruses. Intervirology. 1982;18(4):177–188. doi: 10.1159/000149323. [DOI] [PubMed] [Google Scholar]
  19. Nowak J. A., Das J., Maniloff J. Characterization of an Acholeplasma laidlawii variant with a REP- phenotype. J Bacteriol. 1976 Aug;127(2):832–836. doi: 10.1128/jb.127.2.832-836.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nowak J. A., Maniloff J. Physical characterization of the superhelical DNA genome of an enveloped mycoplasmavirus. J Virol. 1979 Jan;29(1):374–380. doi: 10.1128/jvi.29.1.374-380.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Scott J. F., Eisenberg S., Bertsch L. L., Kornberg A. A mechanism of duplex DNA replication revealed by enzymatic studies of phage phi X174: catalytic strand separation in advance of replication. Proc Natl Acad Sci U S A. 1977 Jan;74(1):193–197. doi: 10.1073/pnas.74.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sladek T. L., Maniloff J. Polyethylene glycol-dependent transfection of Acholeplasma laidlawii with mycoplasma virus L2 DNA. J Bacteriol. 1983 Aug;155(2):734–741. doi: 10.1128/jb.155.2.734-741.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES