Analysis of the Protein-Coding Content of the Sequence of Human Cytomegalovirus Strain AD169

Chee, Mark S.; Bankier, Alan T.; Beck, Stephan; Böhni, Ruth; Brown, C. B.; Cerny, Ronald L.; Horsnell, Terry; Hutchison, Clyde A.; Kouzarides, Tony; Martignetti, John A.; Preddie, E.; Satchwell, Sandra C.; Tomlinson, Peter; Weston, Kathleen; Barrell, Bart

doi:10.1007/978-3-642-74980-3_6

Cited by 950 publications

(1,134 citation statements)

References 169 publications

Supporting

Mentioning

1,108

Contrasting

Unclassified

Order By: Relevance

“…This plasmid contained 12 kB of genomic sequences of HCMV strain Ad169 from nucleotides 115926 to 127969. 29 The GFP-Neo expression cassette was amplified by PCR using the vector pQBI-PGK (Quantum Biotechnologies Inc., Quebec, Canada) as template. As no information about the requirements and mechanisms of DB-formation were available, the GFP-Neo expression cassette was inserted in-frame in the 3 0 -portion of the pp65-gene, using a SrfI site of plasmid pAD6-SpeI.…”

mentioning

confidence: 99%

Protein delivery by subviral particles of human cytomegalovirus

Pepperl-Klindworth¹,

Frankenberg²,

Riegler

et al. 2003

Gene Ther

View full text Add to dashboard Cite

Direct protein delivery is an emerging technology in vaccine development and gene therapy. We could previously show that subviral dense bodies (DB) of human cytomegalovirus (HCMV), a beta-herpesvirus, transport viral proteins into target cells by membrane fusion. Thus these non-infectious particles provide a candidate delivery system for the prophylactic and therapeutic application of proteins. Here we provide proof of principle that DB can be modified genetically. A 55 kDa fusion protein consisting of the green fluorescent protein and the neomycin phosphotransferase could be packed in and delivered into cells by recombinant DB in a functional fashion. Furthermore, transfer of protein into fibroblasts and dendritic cells by DB was efficient, leading to exogenous loading of the MHC-class I antigen presentation pathway. Thus, DB may be a promising basis for the development of novel vaccine strategies and therapeutics based on recombinant polypeptides.

show abstract

mentioning

confidence: 99%

Protein delivery by subviral particles of human cytomegalovirus

Pepperl-Klindworth¹,

Frankenberg²,

Riegler

et al. 2003

Gene Ther

View full text Add to dashboard Cite

show abstract

“…The ORF of the MCMV ICP18.5 gene overlaps the ORF of the glyeoprotein B gene by 32 eodons. The nueleotide sequenees of the MCMV ICP18.5 and MDBP ORFs showed 65 and 60% homology to the UL56 and UL57 ORFs of the HCMV genome (Chee et al, 1990). Remarkably, the sequenees upstream of the MDBP ORF (map units 0.401 to 0.406) and the 200 nt between the MDBP and ICP18.5 ORFs displayalmost no similarity to the eorresponding sequenees in the HCMV genome.…”

Section: Resultsmentioning

confidence: 97%

“…The HCMV strain A0169 MOB protein consists of 1235 amino acids (Chee et al, 1990) and the apparent molecular masses of the MOB proteins of simian (strain Colburn) and human CMV have been determined to 129 and 140 kOa (Anders et al, 1986;Kemble et al, 1987). Thus, the predicted molecular mass of the MCMV MOB protein is in the same range as the observed molecular masses of the SCMV and HCMV MOB proteins.…”

Section: Discussionmentioning

confidence: 99%

“…However, a molecular approach to virus control will only be feasible when the factors contributing to viral pathogenesis and antiviral immunity are defined at the genetic level. Whereas the complete sequence of the HCMV genome has been determined (Chee et a/., 1990), only a few MCMV genes and proteins have been analyzed so far (Keil et a/., 1987;Bühler et a/., 1990;Elliott et a/., 1991;Loh et a/., 1991;Messerle et a/., 1992;Rapp et a/., 1992). In order to study the homology of the two CMV genomes we have determined the nucleic acid sequence of a region of the MCMV genome, which contains aseries of genes conserved throughout all herpesviruses.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of the murine cytomegalovirus genes encoding the major DNA binding protein and the ICP18.5 homolog

Messerle

Keil²,

Schneider

et al. 1992

Virology

View full text Add to dashboard Cite

In several herpesviruses the genes for the major DNA binding protein (MDBP), a putative assembly protein, the glycoprotein B (gB), and the viral DNA polymerase (pol) coliocate. In murine cytomegalovirus (MCMV), two members of this gene block, pol (Elliott, Clark, Jaquish, and Spector, 1991, Virology 185, 169-186) and gB (Rapp, Messerle, BOhler, Tannheimer, Keil, and Koszinowski, 1992, J. Virol., 66,4399-4406) have been characterized. Here the two other MCMV genes are characterized, the gene encoding the MDBP and the ICP18.5 homolog encoding a putative assembly protein. Like in human cytomegalovirus (HCMV) the genes order is pol, gB, ICP18.5, and MDBP. The 4.2-kb MDBP mRNA is expressed first in the early phase, whereas the 3.0-kb ICP18.5 mRNA is a late transcript. The open reading frame of the MDBP gene has the capacity of encoding a protein of 1191 amino acids with a predicted molecular mass of 131.7 kDa. The MCMV ICP18.5 ORF is translated into a polypeptide of 798 amino acids with a calculated molecular mass of 89.1 kDa. Comparison of the amino acid sequences of the predicted proteins of MCMV with the respective proteins of HCMV, Epstein-Barr virus (EBV), and herpes simplex virus type-1 (HSV-1) reveals a striking homology ranging from 72% (HCMV), 50% (EBV), to 45% (HSV-1) for the MDBP sequence and from 74% (HCMV), 51 % (EBV), to 49% (HSV-1) for the ICP18.5 sequence. These results establish the elose relationship of the two cytomegaloviruses, and underline the usefulness of the murine model for studies on the biology of the CMV infection.

show abstract

“…Additionally, some are encoded by virus genes. To date, 20 putative viral GCR genes have been discovered: 2 in poxvirus genomes (21,44), 11 in betaherpesvirus genomes (9,22,33,49,54), and 7 in gammaherpesvirus genomes (4,26,48,57,63). The majority of these genes was found to be similar to genes encoding cellular chemokine receptors.…”

mentioning

confidence: 99%

Deletion of the R78 G protein-coupled receptor gene from rat cytomegalovirus results in an attenuated, syncytium-inducing mutant strain

Vink¹

1999

Journal of Clinical Virology

View full text Add to dashboard Cite

The rat cytomegalovirus (RCMV) R78 gene belongs to an uncharacterized class of viral G protein-coupled receptor (GCR) genes. The predicted amino acid sequence of the R78 open reading frame (ORF) shows 25 and 20% similarity with the gene products of murine cytomegalovirus M78 and human cytomegalovirus UL78, respectively. The R78 gene is transcribed throughout the early and late phases of infection in rat embryo fibroblasts (REF) in vitro. Transcription of R78 was found to result in three different mRNAs: (i) a 1.8-kb mRNA containing the R78 sequence, (ii) a 3.7-kb mRNA containing both R77 and R78 sequences, and (iii) a 5.7-kb mRNA containing at least ORF R77 and ORF R78 sequences. To investigate the function of the R78 gene, we generated two different recombinant virus strains: an RCMV R78 null mutant (RCMV⌬R78a) and an RCMV mutant encoding a GCR from which the putative intracellular C terminus has been deleted (RCMV⌬R78c). These recombinant viruses replicated with a 10-to 100-fold-lower efficiency than wild-type (wt) virus in vitro. Interestingly, unlike wt virus-infected REF, REF infected with the recombinants develop a syncytium-like appearance. A striking difference between wt and recombinant viruses was also seen in vivo: a considerably higher survival was seen among recombinant virus-infected rats than among RCMV-infected rats. We conclude that the RCMV R78 gene encodes a novel GCR-like polypeptide that plays an important role in both RCMV replication in vitro and the pathogenesis of viral infection in vivo.

show abstract

Analysis of the Protein-Coding Content of the Sequence of Human Cytomegalovirus Strain AD169

Cited by 950 publications

References 169 publications

Protein delivery by subviral particles of human cytomegalovirus

Protein delivery by subviral particles of human cytomegalovirus

Characterization of the murine cytomegalovirus genes encoding the major DNA binding protein and the ICP18.5 homolog

Deletion of the R78 G protein-coupled receptor gene from rat cytomegalovirus results in an attenuated, syncytium-inducing mutant strain

Contact Info

Product

Resources

About