1887

Abstract

The lytic-phase origin of DNA replication from human herpesvirus 6B (HHV-6B Lyt) contains two binding sites for the origin-binding protein (OBP), both of which are required for DNA replication and which are separated by an AT-rich spacer. We have tested the functional significance of the structural, spatial and sequence characteristics of this spacer element by constructing a series of mutated origin sequences and analysing their replication efficiency. Changes in the sequence composition or length of the spacer resulted in dramatic decreases in replication efficiency. Furthermore, in contrast to what has been observed for herpes simplex virus type 1 (HSV-1) ori, insertion of a complete helical turn of DNA into the spacer also resulted in abrogation of origin function. These data suggest that the arrangement of OBP sites in HHV-6B oriLyt is stringently constrained in terms of spacing and intervening sequence.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-78-5-1125
1997-05-01
2024-11-04
Loading full text...

Full text loading...

/deliver/fulltext/jgv/78/5/9152433.html?itemId=/content/journal/jgv/10.1099/0022-1317-78-5-1125&mimeType=html&fmt=ahah

References

  1. Boehmer P. E., Craigie M. C., Stow N. D., Lehman I. R. 1994; Association of origin binding protein and single strand DNA-binding protein, ICP8, during herpes simplex virus type 1 DNA replication in vivo. Journal of Biological Chemistry 269:29329–29334
    [Google Scholar]
  2. Dabrowski C. E., Carmillo P. J., Schaffer P. A. 1994; Cellular protein interactions with herpes simplex virus type 1 oriS. Moleculan and Cellular Biology 14:2545–2555
    [Google Scholar]
  3. Dewhurst S., Dollard S. C., Pellett P. E., Dambaugh T. R. 1993; Identification of a lytic-phase origin of DNA replication in human herpesvirus 6B strain Z29. Journal of Virology 67:7680–7683
    [Google Scholar]
  4. Dewhurst S., Krenitsky D. M., Dykes C. 1994; Human herpesvirus 6B origin: sequence diversity, requirement for two binding sites for origin-binding protein, and enhanced replication from origin multimers. Journal of Virology 68:6799–6803
    [Google Scholar]
  5. Elias P., Gustafsson C. M., Hammarsten O. 1990; The origin binding protein of herpes simplex virus 1 binds cooperatively to the viral origin of replication oris. Journal of Biological Chemistry 265:17167–17173
    [Google Scholar]
  6. Fierer D. S., Challberg M. D. 1995; The stoichiometry of binding of the herpes simplex virus type 1 origin binding protein, UL9, to OriS. Journal of Biological Chemistry 270:7330–7334
    [Google Scholar]
  7. Gruffat H., Renner O., Pich D., Hammerschmidt W. 1995; Cellular proteins bind to the downstream component of the lytic origin of DNA replication of Epstein-Barr virus. Journal of Virology 69:1878–1886
    [Google Scholar]
  8. Gustafsson C. M., Hammarsten O., Falkenberg M., Elias P. 1994; Herpes simplex virus DNA replication: a spacer sequence directs the ATP-dependent formation of a nucleoprotein complex at oriS. Proceedings of the National Academy of Sciences, USA 91:4629–4633
    [Google Scholar]
  9. Inoue N., Pellett P. E. 1995; Human herpesvirus 6B origin-binding protein: DNA-binding domain and consensus binding sequence. Journal of Virology 69:4619–4627
    [Google Scholar]
  10. Inoue N., Dambaugh T. R., Rapp J. C., Pellett P. E. 1994; Alphaherpesvirus origin-binding protein homolog encoded by human herpesvirus 6B, a betaherpesvirus, binds to nucleotide sequences that are similar to ori regions of alphaherpesviruses. Journal of Virology 68:4126–4136
    [Google Scholar]
  11. Koff A., Schwedes J. F., Tegtmeyer P. 1991; Herpes simplex virus origin-binding protein (UL9) loops and distorts the viral replication origin. Journal of Virology 65:3284–3292
    [Google Scholar]
  12. Lockshon D., Galloway D. A. 1986; Cloning and characterization of oriL2, a large palindromic DNA replication origin of herpes simplex virus type 2. Journal of Virology 58:513–521
    [Google Scholar]
  13. Lockshon D., Galloway D. A. 1988; Sequence and structural requirements of a herpes simplex viral DNA replication origin. Molecular and Cellular Biology 8:4018–4027
    [Google Scholar]
  14. Martin D. W., Deb S. P., Klauer J. S., Deb S. 1991; Analysis of the herpes simplex virus type 1 OriS sequence : mapping of functional domains. Journal of Virology 65:4359–4369
    [Google Scholar]
  15. McLean G. W., Abbotts A. P., Parry M. E., Marsden H. S., Stow N. D. 1994; The herpes simplex virus type 1 origin-binding protein interacts specifically with the viral UL8 protein. Journal ofGeneral Virology 75:2699–2706
    [Google Scholar]
  16. Stow N. D., McMonagle E. C. 1983; Characterization of the TRS/IRS origin of DNA replication of herpes simplex virus type 1. Virology 130:427–438
    [Google Scholar]
  17. Weller S. K., Spadaro A., Schaffer J. E., Murray A. W., Maxam A. M., Schaffer P. A. 1985; Cloning, sequencing, and functional analysis of oriL, a herpes simplex virus type 1 origin of DNA synthesis. Molecular and Cellular Biology 5:930–942
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-78-5-1125
Loading
/content/journal/jgv/10.1099/0022-1317-78-5-1125
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error