A mass spectrometry-based proteomic approach to study Marek's Disease Virus gene expression.
Marek's Disease Virus (MDV) is an avian herpesvirus that causes a lymphoproliferative disorder in chickens. MDV transitions between a lytic phase in which new viruses are produced and a latent phase in which the virus lays dormant. The mechanism controlling this lytic-to-latent switch remains unclear. To better understand the lytic phase of MDV infection, a mass spectrometry-based strategy was developed to identify viral proteins and to qualitatively examine their abundance in lytically infected chicken embryo fibroblast (CEF) cells. A combination of strong cation exchange chromatography (SCXC) and microcapillary reversed-phase liquid chromatography-tandem mass spectrometry (murpLC/MS/MS) was used to resolve peptides from tryptic digests of MDV-infected CEF cell lysates. Peptides were identified by searching the tandem mass spectra against a protein database containing both MDV proteins and all currently available Gallus gallus proteins using the SEQUEST algorithm. A total of 427 MDV peptides, corresponding to 82 unique proteins, were identified, with 56 of them detected with at least two unique peptides. Overall, nearly 80% of all putative MDV proteins expressed in infected CEF cells were identified. We anticipate that this approach will be a viable method for determining how viral and host proteome changes occurring in Marek's Disease pathogenesis regulate the switch between the lytic and latent phases of the MDV life cycle.
Department of Animal Science, North Carolina State University, Raleigh, NC 27695-7621, USA.
Hsiao-Ching S
HC
Liu
Erik J
EJ
Soderblom
Michael B
MB
Goshe
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002852:Chromatography, Ion Exchange; D019295:Computational Biology; D030562:Databases, Protein; D005347:Fibroblasts / Q000821:virology; D008401:Gas Chromatography-Mass Spectrometry; D020869:Gene Expression Profiling / Q000379:methods*; D008381:Herpesvirus 2, Gallid / Q000235:genetics*; D020543:Proteome / Q000032:analysis / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods; D014357:Trypsin / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification
D020543:Proteome; D014764:Viral Proteins; D014357:Trypsin