AI Article Synopsis
- Researchers identified proteins in murine cytomegalovirus (MCMV) using both proteomic and genomic techniques, analyzing purified viral particles through processes like gel electrophoresis and mass spectrometry.
- A total of 38 proteins from various viral families were identified, along with 20 genes of unknown function, while some coding irregularities were found in specific protein sequences and confirmed through further sequencing.
- Additionally, mass spectrometry suggested new unannotated ORFs, and immunoblot tests verified the expression of one of these ORFs, m166.5, during viral infection.
Article Abstract
Proteins associated with the murine cytomegalovirus (MCMV) viral particle were identified by a combined approach of proteomic and genomic methods. Purified MCMV virions were dissociated by complete denaturation and subjected to either separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and in-gel digestion or treated directly by in-solution tryptic digestion. Peptides were separated by nanoflow liquid chromatography and analyzed by tandem mass spectrometry (LC-MS/MS). The MS/MS spectra obtained were searched against a database of MCMV open reading frames (ORFs) predicted to be protein coding by an MCMV-specific version of the gene prediction algorithm GeneMarkS. We identified 38 proteins from the capsid, tegument, glycoprotein, replication, and immunomodulatory protein families, as well as 20 genes of unknown function. Observed irregularities in coding potential suggested possible sequence errors in the 3'-proximal ends of m20 and M31. These errors were experimentally confirmed by sequencing analysis. The MS data further indicated the presence of peptides derived from the unannotated ORFs ORF(c225441-226898) (m166.5) and ORF(105932-106072). Immunoblot experiments confirmed expression of m166.5 during viral infection.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC521832 | PMC |
| http://dx.doi.org/10.1128/JVI.78.20.11187-11197.2004 | DOI Listing |
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