Poxvirus Proteomics and Virus-Host Protein Interactions

Abstract

SUMMARY Studies of the functional proteins encoded by the poxvirus genome provide information about the composition of the virus as well as individual virus-virus protein and virus-host protein interactions, which provides insight into viral pathogenesis and drug discovery. Widely used proteomic techniques to identify and characterize specific protein-protein interactions include yeast two-hybrid studies and coimmunoprecipitations. Recently, various mass spectrometry techniques have been employed to identify viral protein components of larger complexes. These methods, combined with structural studies, can provide new information about the putative functions of viral proteins as well as insights into virus-host interaction dynamics. For viral proteins of unknown function, identification of either viral or host binding partners provides clues about their putative function. In this review, we discuss poxvirus proteomics, including the use of proteomic methodologies to identify viral components and virus-host protein interactions. High-throughput global protein expression studies using protein chip technology as well as new methods for validating putative protein-protein interactions are also discussed.

Generation of a Complete Single-Gene Knockout Bacterial Artificial Chromosome Library of Cowpox Virus and Identification of Its Essential Genes

“…In accordance with previous studies, we found that 71% of all late genes were essential for virus replication. The products of all essential late genes are predicted to be incorporated into mature virions (24)(25)(26)31). We also confirmed that most essential genes were located in the central portion of the genome, whereas most nonessential genes clustered toward the genomic termini.…”

“…Searches for homologous poxvirus sequences, as well as prediction of gene expression kinetics and function of encoded proteins, were performed using the NCBI BLAST and the VectorNTI 9.1 software package (Invitrogen, Darmstadt, Germany) and were based on OPV sequences available at the Poxvirus Bioinformatics Resource Center (PBRC; http://www.poxvirus.org) and GenBank (23). Predicted incorporation of CPXV proteins into the virion was based on information for homologous proteins of VACV (7,(24)(25)(26).…”

Human CD4+T Cell Response to Human Herpesvirus 6

“…HHV-6A and HHV-6B variants share a high degree of homology, reaching up to 99% in central regions of the genome that contain the core genes, but with less conservation and even variant-specific genes in the flanking regions (73). Epidemiological studies suggest that the HHV-6B variant is widely distributed in the world, infecting over 90% of people (46).…”

Cell- and Virus-Mediated Regulation of the Barrier-to-Autointegration Factor's Phosphorylation State Controls Its DNA Binding, Dimerization, Subcellular Localization, and Antipoxviral Activity

“…Vaccinia virus, the prototypical member of this family, exhibits significant autonomy from the host cell, as demonstrated by its ability to complete its entire life cycle in the cytoplasm of infected cells. Such autonomy is attributed to the large poxviral genome, which encodes essential replication and transcription proteins (18,19). One of these essential proteins is the B1 Ser/Thr kinase, which is expressed early in the viral life cycle and performs functions in viral DNA replication and intermediate gene expression (20)(21)(22)(23)(24)(25)(26).…”