Skip to content
1887

Journal of General Virology header logo Journal of General Virology

Volume 94, Issue 11

Characterization of protein–protein interaction domains within the baculovirus Autographa californica multiple nucleopolyhedrovirus late expression factor LEF-3 No Access

Abstract

Autographa californica nucleopolyhedrovirus late expression factor 3 (LEF-3) is required for late viral gene expression probably through its numerous functions related to DNA replication, including nuclear localization of the virus helicase P143 and binding to ssDNA. LEF-3 appears to interact with itself as a homo-oligomer, although the details of this oligomeric structure are not yet known. To examine LEF-3–LEF-3 interactions, a bimolecular fluorescent protein complementation assay was used. Pairs of recombinant plasmids expressing full-length LEF-3 fused to one of two complementary fragments (V1 or V2) of a variant of yellow fluorescent protein named ‘Venus’ were constructed. Plasmids expressing fusions with complementary fragments of Venus were co-transfected into Sf21 cells and analysed by fluorescence microscopy. Co-transfected plasmids expressing full-length V1–LEF-3 and V2–LEF-3 showed positive fluorescence, confirming the formation of homo-oligomers. A series of truncated V1/V2–LEF-3 fusions was constructed and used to investigate interactions with one another as well as with full-length LEF-3.

  • Received:
  • Accepted:
  • Published Online:
© 2013 SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.056267-0
2013-11-01
2025-05-13
Loading full text...

Full text loading...

References

  1. Ahrens C. H., Carlson C., Rohrmann G. F. 1995; Identification, sequence, and transcriptional analysis of lef-3, a gene essential for Orgyia pseudotsugata baculovirus DNA replication. Virology 210:372–382 [View Article][PubMed]
     [Google Scholar]
  2. Au V., Yu M., Carstens E. B. 2009; Characterization of a baculovirus nuclear localization signal domain in the late expression factor 3 protein. Virology 385:209–217 [View Article][PubMed]
     [Google Scholar]
  3. Chen Z., Carstens E. B. 2005; Identification of domains in Autographa californica multiple nucleopolyhedrovirus late expression factor 3 required for nuclear transport of P143. J Virol 79:10915–10922 [View Article][PubMed]
     [Google Scholar]
  4. Chen T., Sahri D., Carstens E. B. 2004; Characterization of the interaction between P143 and LEF-3 from two different baculovirus species: Choristoneura fumiferana nucleopolyhedrovirus LEF-3 can complement Autographa californica nucleopolyhedrovirus LEF-3 in supporting DNA replication. J Virol 78:329–339 [View Article][PubMed]
     [Google Scholar]
  5. Evans J. T., Rohrmann G. F. 1997; The baculovirus single-stranded DNA binding protein, LEF-3, forms a homotrimer in solution. J Virol 71:3574–3579[PubMed]
     [Google Scholar]
  6. Evans J. T., Rosenblatt G. S., Leisy D. J., Rohrmann G. F. 1999; Characterization of the interaction between the baculovirus ssDNA-binding protein (LEF-3) and putative helicase (P143). J Gen Virol 80:493–500[PubMed]
     [Google Scholar]
  7. Hang X., Dong W., Guarino L. A. 1995; The lef-3 gene of Autographa californica nuclear polyhedrosis virus encodes a single-stranded DNA-binding protein. J Virol 69:3924–3928[PubMed]
     [Google Scholar]
  8. Ito E., Sahri D., Knippers R., Carstens E. B. 2004; Baculovirus proteins IE-1, LEF-3, and P143 interact with DNA in vivo: a formaldehyde cross-linking study. Virology 329:337–347 [View Article][PubMed]
     [Google Scholar]
  9. King A. M. Q., Adams M. J., Carstens E. B., Lefkowitz E. J. 2011 Virus Taxonomy. Ninth Report of the International Committee on Taxonomy of Viruses London, San Diego: Elsevier Academic Press;
     [Google Scholar]
  10. Li Y., Passarelli A. L., Miller L. K. 1993; Identification, sequence, and transcriptional mapping of lef-3, a baculovirus gene involved in late and very late gene expression. J Virol 67:5260–5268[PubMed]
     [Google Scholar]
  11. 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 [View Article][PubMed]
     [Google Scholar]
  12. Mikhailov V. S., Okano K., Rohrmann G. F. 2006; Structural and functional analysis of the baculovirus single-stranded DNA-binding protein LEF-3. Virology 346:469–478 [View Article][PubMed]
     [Google Scholar]
  13. Morell M., Ventura S., Avilés F. X. 2009; Protein complementation assays: approaches for the in vivo analysis of protein interactions. FEBS Lett 583:1684–1691 [View Article][PubMed]
     [Google Scholar]
  14. Nyfeler B., Michnick S. W., Hauri H. P. 2005; Capturing protein interactions in the secretory pathway of living cells. Proc Natl Acad Sci U S A 102:6350–6355 [View Article][PubMed]
     [Google Scholar]
  15. Okano K., Mikhailov V. S., Maeda S. 1999; Colocalization of baculovirus IE-1 and two DNA-binding proteins, DBP and LEF-3, to viral replication factories. J Virol 73:110–119[PubMed]
     [Google Scholar]
  16. Remy I., Michnick S. W. 2007; Application of protein-fragment complementation assays in cell biology. Biotechniques 42:137–145 [View Article][PubMed]
     [Google Scholar]
  17. Rohrmann G. F. 2011 Baculovirus Molecular Biology, 2nd edn. Bethsda (MD): National Library of Medicine (US), NCBI;
     [Google Scholar]
  18. Shereda R. D., Kozlov A. G., Lohman T. M., Cox M. M., Keck J. L. 2008; SSB as an organizer/mobilizer of genome maintenance complexes. Crit Rev Biochem Mol Biol 43:289–318 [View Article][PubMed]
     [Google Scholar]
  19. Shyu Y. J., Liu H., Deng X., Hu C. D. 2006; Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. Biotechniques 40:61–66 [View Article][PubMed]
     [Google Scholar]
  20. Wu Y., Carstens E. B. 1998; A baculovirus single-stranded DNA binding protein, LEF-3, mediates the nuclear localization of the putative helicase P143. Virology 247:32–40 [View Article][PubMed]
     [Google Scholar]
  21. Yu M., Carstens E. B. 2010; Identification of a domain of the baculovirus Autographa californica multiple nucleopolyhedrovirus single-strand DNA-binding protein LEF-3 essential for viral DNA replication. J Virol 84:6153–6162 [View Article][PubMed]
     [Google Scholar]
/content/journal/jgv/10.1099/vir.0.056267-0
Loading
Characterization of protein–protein interaction domains within the baculovirus Autographa californica multiple nucleopolyhedrovirus late expression factor LEF-3
J. Gen. Virol. 94, 2530 (2013); https://doi.org/10.1099/vir.0.056267-0
/content/journal/jgv/10.1099/vir.0.056267-0
/content/journal/jgv/10.1099/vir.0.056267-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

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