multicapsid nucleopolyhedrovirus - gene encodes a functional 3′ to 5′ exonuclease Free

Abstract

The iral hree-prime epair onuclease (-) gene of the multicapsid nucleopolyhedrovirus (AgMNPV) is the first baculovirus gene to be described with significant homology to a 3′ exonuclease. - is an early gene that is expressed by AgMNPV from 3 h post-infection. In the present study, the AgMNPV - ORF was cloned into the baculovirus multicapsid nucleopolyhedrovirus (AcMNPV) under the control of a polyhedrin promoter. The resulting virus produced an abundant, soluble protein that migrated with an apparent size of 23·7 kDa. The 3′ to 5′ exonuclease activity associated with this --expressing recombinant AcMNPV was 2000-fold above that of wild-type AcMNPV. This exonuclease activity was inhibited by EDTA and was activated in the presence of Mg and, to a lesser extent, Mn. From these results, the AgMNPV - gene is concluded to encode an independently active 3′ to 5′ exonuclease.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80109-0
2004-10-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/10/vir852863.html?itemId=/content/journal/jgv/10.1099/vir.0.80109-0&mimeType=html&fmt=ahah

References

  1. Hawtin R. E., Arnold K., Ayres M. D. 7 other authors 1995; Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212:673–685 [CrossRef]
    [Google Scholar]
  2. Hink W. F. 1970; Established insect cell line from the cabbage looper, Trichoplusia ni . Nature 226:466–467
    [Google Scholar]
  3. Kitts P. A., Possee R. D. 1993; A method for producing recombinant baculovirus expression vectors at high frequency. Biotechniques 14:810–817
    [Google Scholar]
  4. Kogan P. H., Blissard G. W. 1994; A baculovirus gp64 early promoter is activated by host transcription factor binding to CACGTG and GATA elements. J Virol 68:813–822
    [Google Scholar]
  5. Li L., Rohrmann G. F. 2000; Characterization of a baculovirus alkaline nuclease. J Virol 74:6401–6407 [CrossRef]
    [Google Scholar]
  6. Mazur D. J., Perrino F. W. 1999; Identification and expression of TREX1 and TREX2 cDNA sequences encoding mammalian 3′→5′ exonucleases. J Biol Chem 274:19655–19660 [CrossRef]
    [Google Scholar]
  7. Mazur D. J., Perrino F. W. 2001; Excision of 3′ termini by Trex1 and TREX2 3′→5′ exonucleases. J Biol Chem 276:17022–17029 [CrossRef]
    [Google Scholar]
  8. Mikhailov V. S., Okano K., Rohrmann G. F. 2003; Baculovirus alkaline nuclease possesses a 5′→3′ exonuclease activity and associates with the DNA-binding protein LEF-3. J Virol 77:2436–2444 [CrossRef]
    [Google Scholar]
  9. Mikhailov V. S., Okano K., Rohrmann G. F. 2004; Specificity of the endonuclease activity of the baculovirus alkaline nuclease for single-stranded DNA. J Biol Chem 279:14734–14745 [CrossRef]
    [Google Scholar]
  10. Moscardi F. 1999; Assessment of the application of baculoviruses for control of lepidoptera. Annu Rev Entomol 44:257–289 [CrossRef]
    [Google Scholar]
  11. Serafini D. M., Schellhorn H. E. 1999; Endonuclease III and endonuclease IV protect Escherichia coli from the lethal and mutagenic effects of near-UV irradiation. Can J Microbiol 45:632–637 [CrossRef]
    [Google Scholar]
  12. Shapiro M., Farrar R. R., Domek J. Jr, Javaid I. 2002; Effects of virus concentration and ultraviolet irradiation on the activity of corn earworm and beet armyworm (Lepidoptera: Noctuidae) nucleopolyhedroviruses. J Econ Entomol 95:243–249 [CrossRef]
    [Google Scholar]
  13. Shippam-Brett C. E., Willis L. G., Theilmann D. A. 2001; Analysis of sequences involved in IE2 transactivation of a baculovirus immediate–early gene promoter and identification of a new regulatory motif. Virus Res 75:13–28 [CrossRef]
    [Google Scholar]
  14. Sieburth P. J., Maruniak J. E. 1988; Susceptibility of an established cell line of Anticarsia gemmatalis (Lepidoptera: Noctuidae) to three nuclear polyhedrosis viruses. J Invertebr Pathol 52:453–458 [CrossRef]
    [Google Scholar]
  15. Slack J. M., Kuzio J., Faulkner P. 1995; Characterization of v - cath , a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J Gen Virol 76:1091–1098 [CrossRef]
    [Google Scholar]
  16. Slack J. M., Ribeiro B. M., Lobo de Souza M. 2004; The gp64 locus of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus contains a 3′ repair exonuclease homologue and lacks v - cath and ChiA genes. J Gen Virol 85:211–219 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80109-0
Loading
/content/journal/jgv/10.1099/vir.0.80109-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed