title: Methods to study the nucleocytoplasmic transport of macromolecules with respect to their impact on the regulation of human cytomegalovirus gene expression.
abstract: One defining feature of eukaryotic cells is their compartmentalization into nucleus and cytoplasm which provides sophisticated opportunities for the regulation of gene expression. Accurate subcellular localization is crucial for the effective function of most viral macromolecules, and nuclear translocation is central to the function of herpesviral proteins that are involved in processes such as transcription or DNA replication. Human cytomegalovirus (HCMV) encodes several transactivator proteins which stimulate viral gene expression either on the transcriptional or posttranscriptional level. In this chapter, we focus on nucleocytoplasmic transport mechanisms of either proteins or RNA that are utilized during HCMV infection. We describe commonly used assays to determine the subcellular localization of a protein, its nucleocytoplasmic shuttling activity, its capacity to export unspliced RNA from the nucleus, and its association with RNA in vivo.
authors.affiliation: Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Thomas
authors.forename: Barbara
authors.initials: B
authors.lastname: Zielke
authors.forename: Nina
authors.initials: N
authors.lastname: Reuter
authors.forename: Thomas
authors.initials: T
authors.lastname: Stamminger
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000235:genetics*; D003587:Cytomegalovirus / Q000235:genetics* / Q000472:pathogenicity; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000821:virology; D015967:Gene Expression Regulation, Viral; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins