Document Detail

Analysis of the contributions of the equine herpesvirus 1 glycoprotein gB homolog to virus entry and direct cell-to-cell spread.
MedLine Citation:
PMID:  9018127     Owner:  NLM     Status:  MEDLINE    
Experiments to analyze the functions of the equine herpesvirus 1 (EHV-1) glycoprotein gB were performed. Cell lines which stably expressed either the full-length EHV-1 gB or only the extracellular portion of gB (amino acids 1 to 844) were constructed and were termed TCgBf and TCgB delta, respectively. Using the cell line TCgBf, a gB-negative viral mutant, L11delta gB, was generated by replacing a 2.1-kb BglII-NruI fragment in the EHV-1 strain RacL11 gB with the Escherichia coli LacZ gene. EHV-1 strain RacL11, the modified live vaccine strain RacH, and L11delta gB were used for functional studies. It was shown that: (i) EHV-1 gB is essential for virus growth in vitro since gB-negative L11delta gB exhibited titers of <10 PFU/ml when grown and titrated on noncomplementing cells. (ii) The cell line expressing truncated gB (TCgB delta) did not complement for the growth of L11delta gB, but the RacH virus grew to titers comparable to those of RacL11 in all cell lines tested. Since RacH had amino acids 944-980 of gB replaced by 7 missense amino acids as determined by nucleotide sequence analysis, the extreme carboxyterminus but not a domain between amino acid residues 845 and 943, probably the transmembrane domain, of EHV-1 gB is dispensable for virus growth in cultured cells. (iii) Single infected cells but no plaque formation were observed after infection of noncomplementing cells with L11delta gB, demonstrating the requirement of EHV-1 gB for direct cell-to-cell spread of infection. (iv) The attachment of gB-negative L11delta gB virions to target cells was similar to both phenotypically complemented L11delta gB and parent RacL11 virus. (v) L11delta gB viral titers could be enhanced by using the fusogen polyethylene glycol (PEG). The increase of L11delta gB titers by PEG treatment, however, was considerably lower compared to gB-negative pseudorabies virus, suggesting that EHV-1 gB might not be as stringently required for virus penetration as are its homologs in other Alphaherpesvirinae.
A Neubauer; B Braun; C Brandmuller; O R Kaaden; N Osterrieder
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Virology     Volume:  227     ISSN:  0042-6822     ISO Abbreviation:  Virology     Publication Date:  1997 Jan 
Date Detail:
Created Date:  1997-03-04     Completed Date:  1997-03-04     Revised Date:  2008-08-26    
Medline Journal Info:
Nlm Unique ID:  0110674     Medline TA:  Virology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  281-94     Citation Subset:  IM    
Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilians-University Munich, Germany.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/X95374;  X95377
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MeSH Terms
Amino Acid Sequence
Base Sequence
Cell Line
DNA Primers
Fluorescent Antibody Technique, Indirect
Herpesvirus 1, Equid / genetics,  pathogenicity,  physiology*
Molecular Sequence Data
Polyethylene Glycols
Polymerase Chain Reaction
Recombinant Proteins / biosynthesis,  chemistry
Restriction Mapping
Sequence Homology, Amino Acid
Viral Envelope Proteins / biosynthesis,  chemistry,  physiology*
Virion / physiology
Reg. No./Substance:
0/DNA Primers; 0/Polyethylene Glycols; 0/Recombinant Proteins; 0/Viral Envelope Proteins; 0/glycoprotein B, Simplexvirus

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