Evidence from Cauliflower Mosaic Virus Virion DNA for Additional Discontinuities in the Plus Strand

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1985 Oct 25

PMID 2414738

Citations 1

Authors

A J Maule

C M Thomas

Affiliations

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Abstract

Two-dimensional electrophoresis of cauliflower mosaic virus (CaMV) virion DNA and analysis of Southern blots using (+) strand-specific probes to the 5' termini of the beta (5.4 Kb) and alpha (2.6 Kb) strands, revealed the presence of molecules in addition to those predicted from the known structure of CaMV DNA. The presence of 8 Kb molecules of (+) sense after denaturation suggested that a small proportion of circular molecules have only a single discontinuity in the (+) strand. Other molecules, probably 5' coterminal with the beta strand but smaller than 5.4 Kb, indicated that a minority of the circular full length CaMV DNA contain additional gaps in the (+) strand. Consequently, molecules equivalent to the remainder of the beta strand could be identified using a single strand probe for a region towards the 3'-end of the beta strand. Computer analysis of the nucleotide sequence of CaMV DNA in the region of the proposed additional discontinuities revealed regions displaying some homology with the major (+) strand priming sites at the 5' ends of the beta and alpha strands. It is our contention that the additional (+) strand molecules of beta specificity are a consequence of minor (+) strand priming sites.

Citing Articles

Hairpin DNAs of cauliflower mosaic virus generated by reverse transcription in vivo.

Covey S, Turner D EMBO J. 1986; 5(11):2763-8.

PMID: 16453720 PMC: 1167220. DOI: 10.1002/j.1460-2075.1986.tb04565.x.

References

Rigby P, Dieckmann M, Rhodes C, Berg P . Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977; 113(1):237-51. DOI: 10.1016/0022-2836(77)90052-3. View

Taylor J, Cywinski A, Smith J . Discontinuities in the DNA synthesized by an avian retrovirus. J Virol. 1983; 48(3):654-9. PMC: 255397. DOI: 10.1128/JVI.48.3.654-659.1983. View

Hsu T, Taylor J . Single-stranded regions on unintegrated avian retrovirus DNA. J Virol. 1982; 44(1):47-53. PMC: 256239. DOI: 10.1128/JVI.44.1.47-53.1982. View

BOONE L, Skalka A . Viral DNA synthesized in vitro by avian retrovirus particles permeabilized with melittin. II. Evidence for a strand displacement mechanism in plus-strand synthesis. J Virol. 1981; 37(1):117-26. PMC: 170988. DOI: 10.1128/JVI.37.1.117-126.1981. View

Pfeiffer P, Hohn T . Involvement of reverse transcription in the replication of cauliflower mosaic virus: a detailed model and test of some aspects. Cell. 1983; 33(3):781-9. DOI: 10.1016/0092-8674(83)90020-x. View

Maule A, Hull R, Donson J . The application of spot hybridization to the detection of DNA and RNA viruses in plant tissues. J Virol Methods. 1983; 6(4):215-24. DOI: 10.1016/0166-0934(83)90048-4. View

Gilboa E, Goff S, Shields A, Yoshimura F, Mitra S, Baltimore D . In vitro synthesis of a 9 kbp terminally redundant DNA carrying the infectivity of Moloney murine leukemia virus. Cell. 1979; 16(4):863-74. DOI: 10.1016/0092-8674(79)90101-6. View

Volovitch M, Drugeon C, Yot P . Studies on the single-stranded discontinuities of the cauliflower mosaic virus genome. Nucleic Acids Res. 1978; 5(8):2913-25. PMC: 342215. DOI: 10.1093/nar/5.8.2913. View

Covey S, Turner D, Mulder G . A small DNA molecule containing covalently-linked ribonucleotides originates from the large intergenic region of the cauliflower mosaic virus genome. Nucleic Acids Res. 1983; 11(2):251-64. PMC: 325712. DOI: 10.1093/nar/11.2.251. View

10.

Staden R . Computer methods to locate signals in nucleic acid sequences. Nucleic Acids Res. 1984; 12(1 Pt 2):505-19. PMC: 321067. DOI: 10.1093/nar/12.1part2.505. View

11.

Varmus H, Heasley S, Kung H, Oppermann H, Smith V, Bishop J . Kinetics of synthesis, structure and purification of avian sarcoma virus-specific DNA made in the cytoplasm of acutely infected cells. J Mol Biol. 1978; 120(1):55-82. DOI: 10.1016/0022-2836(78)90295-4. View

12.

Thomas C, Hull R, Bryant J, Maule A . Isolation of a fraction from cauliflower mosaic virus-infected protoplasts which is active in the synthesis of (+) and (-) strand viral DNA and reverse transcription of primed RNA templates. Nucleic Acids Res. 1985; 13(12):4557-76. PMC: 321806. DOI: 10.1093/nar/13.12.4557. View

13.

Brown D, Frampton J, Goelet P, Karn J . Sensitive detection of RNA using strand-specific M13 probes. Gene. 1982; 20(2):139-44. DOI: 10.1016/0378-1119(82)90032-4. View

14.

Messing J . New M13 vectors for cloning. Methods Enzymol. 1983; 101:20-78. DOI: 10.1016/0076-6879(83)01005-8. View

15.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

16.

Franck A, Guilley H, Jonard G, Richards K, Hirth L . Nucleotide sequence of cauliflower mosaic virus DNA. Cell. 1980; 21(1):285-94. DOI: 10.1016/0092-8674(80)90136-1. View

17.

Guilley H, Richards K, Jonard G . Observations concerning the discontinuous DNAs of cauliflower mosaic virus. EMBO J. 1983; 2(2):277-82. PMC: 555125. DOI: 10.1002/j.1460-2075.1983.tb01417.x. View

18.

Favaloro J, Treisman R, Kamen R . Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980; 65(1):718-49. DOI: 10.1016/s0076-6879(80)65070-8. View

19.

Gardner R, Howarth A, Hahn P, Brown-Luedi M, Shepherd R, Messing J . The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Res. 1981; 9(12):2871-88. PMC: 326899. DOI: 10.1093/nar/9.12.2871. View

20.

Varmus H . Form and function of retroviral proviruses. Science. 1982; 216(4548):812-20. DOI: 10.1126/science.6177038. View