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. 1997 Feb;71(2):1405–1416. doi: 10.1128/jvi.71.2.1405-1416.1997

A novel cellular site-specific DNA-binding protein cooperates with the viral NS1 polypeptide to initiate parvovirus DNA replication.

J Christensen 1, S F Cotmore 1, P Tattersall 1
PMCID: PMC191197  PMID: 8995666

Abstract

Replication of linear single-stranded parvovirus DNA proceeds by a rolling-hairpin mechanism which generates long, palindromic, duplex concatamers. Processing to monomer length requires initiation from origins of DNA replication located at the 3' and 5' ends of each embedded monomer, reactions which can be recapitulated in vitro for minute virus of mice (MVM). To determine which cellular proteins were essential for replication from these origins, S100 extracts from 293S cells were fractionated on phosphocellulose. When recombined, these fractions were able to support replication in vitro, dependent on the viral initiator protein NS1, using plasmid forms of the 5' origin or the minimal 3' origin as templates. Fraction P-cell 1 contains two factors, replication protein A (RPA) and proliferating-cell nuclear antigen (PCNA), known to be essential for simian virus 40 replication in vitro. When P-cell 1 was replaced with purified recombinant RPA and PCNA, NS1-mediated MVM replication initiated from the 5' origin but not from the 3' origin. The 3' origin is a 50-bp sequence containing three distinct recognition elements, an NS1 binding site, a site at which NS1 nicks the DNA to generate the priming 3' OH, and a region containing a consensus activated transcription factor (ATF) binding site. To identify the missing factor(s) for 3' origin replication, P-cell 1 was fractionated by further chromatography and active fractions were identified by their ability to complement RPA, PCNA, and P-cell 2 for NS1-mediated, origin-specific replication. Gel shift and UV cross-linking analysis of the replication-competent fractions revealed a novel 110-kDa sequence-specific DNA binding protein which recognized the consensus ATF binding site region of the origin and which we have termed parvovirus initiation factor, or PIF. Binding of PIF appears to activate the endonuclease function of NS1, allowing efficient and specific nicking of the 3' minimal origin under stringent conditions in vitro.

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Selected References

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  1. Astell C. R., Chow M. B., Ward D. C. Sequence analysis of the termini of virion and replicative forms of minute virus of mice DNA suggests a modified rolling hairpin model for autonomous parvovirus DNA replication. J Virol. 1985 Apr;54(1):171–177. doi: 10.1128/jvi.54.1.171-177.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen C. Y., Bessesen D. H., Jackson S. M., Hoeffler J. P. Overexpression and purification of transcriptionally competent CREB from a recombinant baculovirus. Protein Expr Purif. 1991 Oct-Dec;2(5-6):402–411. doi: 10.1016/1046-5928(91)90101-n. [DOI] [PubMed] [Google Scholar]
  3. Chen M., Mermod N., Horwitz M. S. Protein-protein interactions between adenovirus DNA polymerase and nuclear factor I mediate formation of the DNA replication preinitiation complex. J Biol Chem. 1990 Oct 25;265(30):18634–18642. [PubMed] [Google Scholar]
  4. Christensen J., Cotmore S. F., Tattersall P. Minute virus of mice transcriptional activator protein NS1 binds directly to the transactivation region of the viral P38 promoter in a strictly ATP-dependent manner. J Virol. 1995 Sep;69(9):5422–5430. doi: 10.1128/jvi.69.9.5422-5430.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Christensen J., Pedersen M., Aasted B., Alexandersen S. Purification and characterization of the major nonstructural protein (NS-1) of Aleutian mink disease parvovirus. J Virol. 1995 Mar;69(3):1802–1809. doi: 10.1128/jvi.69.3.1802-1809.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christensen J., Storgaard T., Bloch B., Alexandersen S., Aasted B. Expression of Aleutian mink disease parvovirus proteins in a baculovirus vector system. J Virol. 1993 Jan;67(1):229–238. doi: 10.1128/jvi.67.1.229-238.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cotmore S. F., Christensen J., Nüesch J. P., Tattersall P. The NS1 polypeptide of the murine parvovirus minute virus of mice binds to DNA sequences containing the motif [ACCA]2-3. J Virol. 1995 Mar;69(3):1652–1660. doi: 10.1128/jvi.69.3.1652-1660.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cotmore S. F., Nüesch J. P., Tattersall P. Asymmetric resolution of a parvovirus palindrome in vitro. J Virol. 1993 Mar;67(3):1579–1589. doi: 10.1128/jvi.67.3.1579-1589.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cotmore S. F., Tattersall P. An asymmetric nucleotide in the parvoviral 3' hairpin directs segregation of a single active origin of DNA replication. EMBO J. 1994 Sep 1;13(17):4145–4152. doi: 10.1002/j.1460-2075.1994.tb06732.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cotmore S. F., Tattersall P. The NS-1 polypeptide of minute virus of mice is covalently attached to the 5' termini of duplex replicative-form DNA and progeny single strands. J Virol. 1988 Mar;62(3):851–860. doi: 10.1128/jvi.62.3.851-860.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Faisst S., Perros M., Deleu L., Spruyt N., Rommelaere J. Mapping of upstream regulatory elements in the P4 promoter of parvovirus minute virus of mice. Virology. 1994 Jul;202(1):466–470. doi: 10.1006/viro.1994.1363. [DOI] [PubMed] [Google Scholar]
  12. Fien K., Stillman B. Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex. Mol Cell Biol. 1992 Jan;12(1):155–163. doi: 10.1128/mcb.12.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hirsch S., Miskimins W. K. Mitogen induction of nuclear factors that interact with a delayed responsive region of the transferrin receptor gene promoter. Cell Growth Differ. 1995 Jun;6(6):719–726. [PubMed] [Google Scholar]
  14. Hölscher C., Kleinschmidt J. A., Bürkle A. High-level expression of adeno-associated virus (AAV) Rep78 or Rep68 protein is sufficient for infectious-particle formation by a rep-negative AAV mutant. J Virol. 1995 Nov;69(11):6880–6885. doi: 10.1128/jvi.69.11.6880-6885.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Karin M., Smeal T. Control of transcription factors by signal transduction pathways: the beginning of the end. Trends Biochem Sci. 1992 Oct;17(10):418–422. doi: 10.1016/0968-0004(92)90012-x. [DOI] [PubMed] [Google Scholar]
  16. Liu F., Green M. R. A specific member of the ATF transcription factor family can mediate transcription activation by the adenovirus E1a protein. Cell. 1990 Jun 29;61(7):1217–1224. doi: 10.1016/0092-8674(90)90686-9. [DOI] [PubMed] [Google Scholar]
  17. Liu Q., Yong C. B., Astell C. R. In vitro resolution of the dimer bridge of the minute virus of mice (MVM) genome supports the modified rolling hairpin model for MVM replication. Virology. 1994 Jun;201(2):251–262. doi: 10.1006/viro.1994.1290. [DOI] [PubMed] [Google Scholar]
  18. Mul Y. M., Verrijzer C. P., van der Vliet P. C. Transcription factors NFI and NFIII/oct-1 function independently, employing different mechanisms to enhance adenovirus DNA replication. J Virol. 1990 Nov;64(11):5510–5518. doi: 10.1128/jvi.64.11.5510-5518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Naeger L. K., Cater J., Pintel D. J. The small nonstructural protein (NS2) of the parvovirus minute virus of mice is required for efficient DNA replication and infectious virus production in a cell-type-specific manner. J Virol. 1990 Dec;64(12):6166–6175. doi: 10.1128/jvi.64.12.6166-6175.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nagata K., Guggenheimer R. A., Hurwitz J. Adenovirus DNA replication in vitro: synthesis of full-length DNA with purified proteins. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4266–4270. doi: 10.1073/pnas.80.14.4266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nüesch J. P., Cotmore S. F., Tattersall P. Sequence motifs in the replicator protein of parvovirus MVM essential for nicking and covalent attachment to the viral origin: identification of the linking tyrosine. Virology. 1995 May 10;209(1):122–135. doi: 10.1006/viro.1995.1236. [DOI] [PubMed] [Google Scholar]
  22. O'Neill E. A., Fletcher C., Burrow C. R., Heintz N., Roeder R. G., Kelly T. J. Transcription factor OTF-1 is functionally identical to the DNA replication factor NF-III. Science. 1988 Sep 2;241(4870):1210–1213. doi: 10.1126/science.3413485. [DOI] [PubMed] [Google Scholar]
  23. Ono M., Tucker P. W., Capra J. D. Production and characterization of recombinant human Ku antigen. Nucleic Acids Res. 1994 Sep 25;22(19):3918–3924. doi: 10.1093/nar/22.19.3918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Perros M., Deleu L., Vanacker J. M., Kherrouche Z., Spruyt N., Faisst S., Rommelaere J. Upstream CREs participate in the basal activity of minute virus of mice promoter P4 and in its stimulation in ras-transformed cells. J Virol. 1995 Sep;69(9):5506–5515. doi: 10.1128/jvi.69.9.5506-5515.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pruijn G. J., van Miltenburg R. T., Claessens J. A., van der Vliet P. C. Interaction between the octamer-binding protein nuclear factor III and the adenovirus origin of DNA replication. J Virol. 1988 Sep;62(9):3092–3102. doi: 10.1128/jvi.62.9.3092-3102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roberts M. R., Han Y., Fienberg A., Hunihan L., Ruddle F. H. A DNA-binding activity, TRAC, specific for the TRA element of the transferrin receptor gene copurifies with the Ku autoantigen. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6354–6358. doi: 10.1073/pnas.91.14.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Traut W., Fanning E. Sequence-specific interactions between a cellular DNA-binding protein and the simian virus 40 origin of DNA replication. Mol Cell Biol. 1988 Feb;8(2):903–911. doi: 10.1128/mcb.8.2.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsurimoto T., Melendy T., Stillman B. Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin. Nature. 1990 Aug 9;346(6284):534–539. doi: 10.1038/346534a0. [DOI] [PubMed] [Google Scholar]
  29. Tsurimoto T., Stillman B. Purification of a cellular replication factor, RF-C, that is required for coordinated synthesis of leading and lagging strands during simian virus 40 DNA replication in vitro. Mol Cell Biol. 1989 Feb;9(2):609–619. doi: 10.1128/mcb.9.2.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tsurimoto T., Stillman B. Replication factors required for SV40 DNA replication in vitro. I. DNA structure-specific recognition of a primer-template junction by eukaryotic DNA polymerases and their accessory proteins. J Biol Chem. 1991 Jan 25;266(3):1950–1960. [PubMed] [Google Scholar]
  31. Tuteja N., Tuteja R., Ochem A., Taneja P., Huang N. W., Simoncsits A., Susic S., Rahman K., Marusic L., Chen J. Human DNA helicase II: a novel DNA unwinding enzyme identified as the Ku autoantigen. EMBO J. 1994 Oct 17;13(20):4991–5001. doi: 10.1002/j.1460-2075.1994.tb06826.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Umbricht C. B., Erdile L. F., Jabs E. W., Kelly T. J. Cloning, overexpression, and genomic mapping of the 14-kDa subunit of human replication protein A. J Biol Chem. 1993 Mar 25;268(9):6131–6138. [PubMed] [Google Scholar]
  33. Umbricht C. B., Griffin C. A., Hawkins A. L., Grzeschik K. H., O'Connell P., Leach R., Green E. D., Kelly T. J. High-resolution genomic mapping of the three human replication protein A genes (RPA1, RPA2, and RPA3). Genomics. 1994 Mar 15;20(2):249–257. doi: 10.1006/geno.1994.1161. [DOI] [PubMed] [Google Scholar]
  34. Wilson G. M., Jindal H. K., Yeung D. E., Chen W., Astell C. R. Expression of minute virus of mice major nonstructural protein in insect cells: purification and identification of ATPase and helicase activities. Virology. 1991 Nov;185(1):90–98. doi: 10.1016/0042-6822(91)90757-3. [DOI] [PubMed] [Google Scholar]
  35. Wold M. S., Weinberg D. H., Virshup D. M., Li J. J., Kelly T. J. Identification of cellular proteins required for simian virus 40 DNA replication. J Biol Chem. 1989 Feb 15;264(5):2801–2809. [PubMed] [Google Scholar]
  36. Yoshida M., Suzuki T., Fujisawa J., Hirai H. HTLV-1 oncoprotein tax and cellular transcription factors. Curr Top Microbiol Immunol. 1995;193:79–89. doi: 10.1007/978-3-642-78929-8_4. [DOI] [PubMed] [Google Scholar]

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