Document Detail


Bimolecular complementation reveals that glycoproteins gB and gH/gL of herpes simplex virus interact with each other during cell fusion.
MedLine Citation:
PMID:  18003913     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Herpes simplex virus entry into cells requires four glycoproteins, gB, gD, gH, and gL. Binding of gD to one of its receptors triggers steps requiring the core fusion proteins, gB and the gH/gL heterodimer. There is evidence that gH/gL initiates hemifusion of cells, but whether this complex interacts physically with gB to cause complete fusion is unknown. We used bimolecular complementation (BiMC) of enhanced yellow fluorescent protein (EYFP) to detect glycoprotein interactions during cell-cell fusion. The N- or C-terminal half of EYFP was fused to the C terminus of gD, gB, and gH to form six chimeric proteins (Dn, Dc, Bn, Bc, Hn, and Hc). BiMC was detected by confocal microscopy. Receptor-bearing (C10) cells cotransfected with Dn and Bc or Dn, Hc, and untagged gL exhibited EYFP fluorescence, indicative of interactions between gD and gB and between gD and gH/gL. EYFP complementation did not occur in cells transfected with gL, Bc, and Hn. However, when gD was coexpressed with these other three proteins, cell-cell fusion occurred and the syncytia exhibited bright EYFP fluorescence. To separate glycoprotein expression from fusion, we transfected C10 cells with gL, Bc, and Hn for 20 h and then added soluble gD to trigger fusion. We detected fluorescent syncytia within 10 min, and both their number and size increased with exposure time to gD. Thus, when gD binds its receptor, the core fusion machinery is triggered to form a multiprotein complex as a step in fusion and possibly virus entry.
Authors:
Doina Atanasiu; J Charles Whitbeck; Tina M Cairns; Brigid Reilly; Gary H Cohen; Roselyn J Eisenberg
Related Documents :
8374773 - Effects of interferons and hydrogen peroxide on ca3 pyramidal cells in rat hippocampal ...
20003813 - Coculturing embryonic stem cells with damaged hepatocytes leads to restoration of damag...
922843 - Unit gravity sedimentation separation of cells comprising the caput epididymidis of the...
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2007-11-14
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  104     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2007 Nov 
Date Detail:
Created Date:  2007-11-27     Completed Date:  2008-01-02     Revised Date:  2013-06-06    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  18718-23     Citation Subset:  IM    
Affiliation:
Department of Microbiology, School of Dental Medicine, and Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line
Cricetinae
Genes, Reporter / genetics
Glycoproteins / genetics,  metabolism*
Membrane Fusion*
Mice
Protein Binding
Simplexvirus / genetics,  metabolism*
Solubility
Viral Proteins / genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
AI-076231/AI/NIAID NIH HHS; AI-18289/AI/NIAID NIH HHS; NS-036731/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Glycoproteins; 0/Viral Proteins
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Mapping hydration dynamics around a protein surface.
Next Document:  Identification of JmjC domain-containing UTX and JMJD3 as histone H3 lysine 27 demethylases.