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Journal of General Virology header logo

Volume 94, Issue 11

Characterization of protein–protein interaction domains within the baculovirus Autographa californica multiple nucleopolyhedrovirus late expression factor LEF-3 Free

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:
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© 2013 SGM
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2013-11-01
2023-03-27
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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]
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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
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