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. 1992 May;4(5):597–608. doi: 10.1105/tpc.4.5.597

A geminivirus replication protein is a sequence-specific DNA binding protein.

E P Fontes 1, V A Luckow 1, L Hanley-Bowdoin 1
PMCID: PMC160156  PMID: 1498611

Abstract

The genome of the geminivirus tomato golden mosaic virus (TGMV) consists of two circular DNA molecules designated as components A and B. The A component encodes the only viral protein, AL1, that is required for viral replication. We showed that AL1 interacts specifically with TGMV A and B DNA by using an immunoprecipitation assay for AL1:DNA complex formation. In this assay, a monoclonal antibody against AL1 precipitated AL1:TGMV DNA complexes, whereas an unrelated antibody failed to precipitate the complexes. Competition assays with homologous and heterologous DNAs established the specificity of AL1:DNA binding. AL1 produced by transgenic tobacco plants and by baculovirus-infected insect cells exhibited similar DNA binding activity. The AL1 binding site maps to 52 bp on the left side of the common region, a 235-bp region that is highly conserved between the two TGMV genome components. The AL1:DNA binding site does not include the putative hairpin structure that is conserved in the common regions or the equivalent 5' intergenic regions of all geminiviruses. These studies demonstrate that a geminivirus replication protein is a sequence-specific DNA binding protein, and the studies have important implications for the role of this protein in virus replication.

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Selected References

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

  1. Accotto G. P., Donson J., Mullineaux P. M. Mapping of Digitaria streak virus transcripts reveals different RNA species from the same transcription unit. EMBO J. 1989 Apr;8(4):1033–1039. doi: 10.1002/j.1460-2075.1989.tb03470.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrews D. L., Beames B., Summers M. D., Park W. D. Characterization of the lipid acyl hydrolase activity of the major potato (Solanum tuberosum) tuber protein, patatin, by cloning and abundant expression in a baculovirus vector. Biochem J. 1988 May 15;252(1):199–206. doi: 10.1042/bj2520199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Billings P. B., Allen R. W., Jensen F. C., Hoch S. O. Anti-RNP monoclonal antibodies derived from a mouse strain with lupus-like autoimmunity. J Immunol. 1982 Mar;128(3):1176–1180. [PubMed] [Google Scholar]
  4. Bisaro D. M., Hamilton W. D., Coutts R. H., Buck K. W. Molecular cloning and characterisation of the two DNA components of tomato golden mosaic virus. Nucleic Acids Res. 1982 Aug 25;10(16):4913–4922. doi: 10.1093/nar/10.16.4913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bronstein I., Edwards B., Voyta J. C. 1,2-dioxetanes: novel chemiluminescent enzyme substrates. Applications to immunoassays. J Biolumin Chemilumin. 1989 Jul;4(1):99–111. doi: 10.1002/bio.1170040116. [DOI] [PubMed] [Google Scholar]
  6. Colasanti J., Tyers M., Sundaresan V. Isolation and characterization of cDNA clones encoding a functional p34cdc2 homologue from Zea mays. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3377–3381. doi: 10.1073/pnas.88.8.3377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Davies J. W., Stanley J. Geminivirus genes and vectors. Trends Genet. 1989 Mar;5(3):77–81. doi: 10.1016/0168-9525(89)90030-9. [DOI] [PubMed] [Google Scholar]
  8. DePamphilis M. L. Transcriptional elements as components of eukaryotic origins of DNA replication. Cell. 1988 Mar 11;52(5):635–638. doi: 10.1016/0092-8674(88)90398-4. [DOI] [PubMed] [Google Scholar]
  9. Eisenberg S., Kornberg A. Purification and characterization of phiX174 gene A protein. A multifunctional enzyme of duplex DNA replication. J Biol Chem. 1979 Jun 25;254(12):5328–5332. [PubMed] [Google Scholar]
  10. Elmer J. S., Brand L., Sunter G., Gardiner W. E., Bisaro D. M., Rogers S. G. Genetic analysis of the tomato golden mosaic virus. II. The product of the AL1 coding sequence is required for replication. Nucleic Acids Res. 1988 Jul 25;16(14B):7043–7060. doi: 10.1093/nar/16.14.7043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Etessami P., Saunders K., Watts J., Stanley J. Mutational analysis of complementary-sense genes of African cassava mosaic virus DNA A. J Gen Virol. 1991 May;72(Pt 5):1005–1012. doi: 10.1099/0022-1317-72-5-1005. [DOI] [PubMed] [Google Scholar]
  12. Feiler H. S., Jacobs T. W. Cell division in higher plants: a cdc2 gene, its 34-kDa product, and histone H1 kinase activity in pea. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5397–5401. doi: 10.1073/pnas.87.14.5397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ferreira P. C., Hemerly A. S., Villarroel R., Van Montagu M., Inzé D. The Arabidopsis functional homolog of the p34cdc2 protein kinase. Plant Cell. 1991 May;3(5):531–540. doi: 10.1105/tpc.3.5.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. GRACE T. D. Establishment of four strains of cells from insect tissues grown in vitro. Nature. 1962 Aug 25;195:788–789. doi: 10.1038/195788a0. [DOI] [PubMed] [Google Scholar]
  15. Gardiner W. E., Sunter G., Brand L., Elmer J. S., Rogers S. G., Bisaro D. M. Genetic analysis of tomato golden mosaic virus: the coat protein is not required for systemic spread or symptom development. EMBO J. 1988 Apr;7(4):899–904. doi: 10.1002/j.1460-2075.1988.tb02894.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hamilton W. D., Bisaro D. M., Coutts R. H., Buck K. W. Demonstration of the bipartite nature of the genome of a single-stranded DNA plant virus by infection with the cloned DNA components. Nucleic Acids Res. 1983 Nov 11;11(21):7387–7396. doi: 10.1093/nar/11.21.7387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hamilton W. D., Stein V. E., Coutts R. H., Buck K. W. Complete nucleotide sequence of the infectious cloned DNA components of tomato golden mosaic virus: potential coding regions and regulatory sequences. EMBO J. 1984 Sep;3(9):2197–2205. doi: 10.1002/j.1460-2075.1984.tb02114.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanley-Bowdoin L., Elmer J. S., Rogers S. G. Expression of functional replication protein from tomato golden mosaic virus in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1446–1450. doi: 10.1073/pnas.87.4.1446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hanley-Bowdoin L., Elmer J. S., Rogers S. G. Transient expression of heterologous RNAs using tomato golden mosaic virus. Nucleic Acids Res. 1988 Nov 25;16(22):10511–10528. doi: 10.1093/nar/16.22.10511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hayes R. J., Buck K. W. Replication of tomato golden mosaic virus DNA B in transgenic plants expressing open reading frames (ORFs) of DNA A: requirement of ORF AL2 for production of single-stranded DNA. Nucleic Acids Res. 1989 Dec 25;17(24):10213–10222. doi: 10.1093/nar/17.24.10213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kunze R., Starlinger P. The putative transposase of transposable element Ac from Zea mays L. interacts with subterminal sequences of Ac. EMBO J. 1989 Nov;8(11):3177–3185. doi: 10.1002/j.1460-2075.1989.tb08476.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Lazarowitz S. G., Pinder A. J., Damsteegt V. D., Rogers S. G. Maize streak virus genes essential for systemic spread and symptom development. EMBO J. 1989 Apr;8(4):1023–1032. doi: 10.1002/j.1460-2075.1989.tb03469.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Luckow V. A., Summers M. D. High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology. 1989 May;170(1):31–39. doi: 10.1016/0042-6822(89)90348-6. [DOI] [PubMed] [Google Scholar]
  26. Luckow V. A., Summers M. D. Signals important for high-level expression of foreign genes in Autographa californica nuclear polyhedrosis virus expression vectors. Virology. 1988 Nov;167(1):56–71. doi: 10.1016/0042-6822(88)90054-2. [DOI] [PubMed] [Google Scholar]
  27. McKay R. D. Binding of a simian virus 40 T antigen-related protein to DNA. J Mol Biol. 1981 Jan 25;145(3):471–488. doi: 10.1016/0022-2836(81)90540-4. [DOI] [PubMed] [Google Scholar]
  28. Meyer T. F., Geider K. Bacteriophage fd gene II-protein. II. Specific cleavage and relaxation of supercoiled RF from filamentous phages. J Biol Chem. 1979 Dec 25;254(24):12642–12646. [PubMed] [Google Scholar]
  29. Revington G. N., Sunter G., Bisaro D. M. DNA sequences essential for replication of the B genome component of tomato golden mosaic virus. Plant Cell. 1989 Oct;1(10):985–992. doi: 10.1105/tpc.1.10.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rogers S. G., Bisaro D. M., Horsch R. B., Fraley R. T., Hoffmann N. L., Brand L., Elmer J. S., Lloyd A. M. Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants. Cell. 1986 May 23;45(4):593–600. doi: 10.1016/0092-8674(86)90291-6. [DOI] [PubMed] [Google Scholar]
  31. Saunders K., Lucy A., Stanley J. DNA forms of the geminivirus African cassava mosaic virus consistent with a rolling circle mechanism of replication. Nucleic Acids Res. 1991 May 11;19(9):2325–2330. doi: 10.1093/nar/19.9.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schalk H. J., Matzeit V., Schiller B., Schell J., Gronenborn B. Wheat dwarf virus, a geminivirus of graminaceous plants needs splicing for replication. EMBO J. 1989 Feb;8(2):359–364. doi: 10.1002/j.1460-2075.1989.tb03385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith G. E., Summers M. D. Restriction Maps of Five Autographa californica MNPV Variants, Trichoplusia ni MNPV, and Galleria mellonella MNPV DNAs with Endonucleases SmaI, KpnI, BamHI, SacI, XhoI, and EcoRI. J Virol. 1979 Jun;30(3):828–838. doi: 10.1128/jvi.30.3.828-838.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stanley J., Frischmuth T., Ellwood S. Defective viral DNA ameliorates symptoms of geminivirus infection in transgenic plants. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6291–6295. doi: 10.1073/pnas.87.16.6291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stenger D. C., Revington G. N., Stevenson M. C., Bisaro D. M. Replicational release of geminivirus genomes from tandemly repeated copies: evidence for rolling-circle replication of a plant viral DNA. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8029–8033. doi: 10.1073/pnas.88.18.8029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sunter G., Bisaro D. M. Transcription map of the B genome component of tomato golden mosaic virus and comparison with A component transcripts. Virology. 1989 Dec;173(2):647–655. doi: 10.1016/0042-6822(89)90577-1. [DOI] [PubMed] [Google Scholar]
  37. Suzuka I., Daidoji H., Matsuoka M., Kadowaki K., Takasaki Y., Nakane P. K., Moriuchi T. Gene for proliferating-cell nuclear antigen (DNA polymerase delta auxiliary protein) is present in both mammalian and higher plant genomes. Proc Natl Acad Sci U S A. 1989 May;86(9):3189–3193. doi: 10.1073/pnas.86.9.3189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]

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