title: Characterization of the interaction between human respiratory syncytial virus and the cell cycle in continuous cell culture and primary human airway epithelial cells.
abstract: Viruses can modify conditions inside cells to make them more favorable for replication and progeny virus production. One way of doing this is through manipulation of the cell cycle, a process that describes the ordered growth and division of cells. Analysis of model cell lines, such as A549 cells and primary airway epithelial cells, infected with human respiratory syncytial virus (HRSV) has shown alteration of the cell cycle during infection, although the signaling events were not clearly understood. In this study, targeted transcriptomic analysis of HRSV-infected primary airway epithelial cells revealed alterations in the abundances of many mRNAs encoding cell cycle-regulatory molecules, including decreases in the D-type cyclins and corresponding cyclin-dependent kinases (CDK4 and CDK6 [CDK4/6]). These alterations were reflected in changes in protein abundance and/or relocalization in HRSV-infected cells; taken together, they were predicted to result in G(0)/G(1) phase arrest. In contrast, there was no change in the abundances of D-type cyclins in A549 cells infected with HRSV. However, the abundance of the G(1)/S phase progression inhibitor p21(WAF1/CIP1) was increased over that in mock-treated cells, and this, again, was predicted to result in G(0)/G(1) phase arrest. The G(0)/G(1) phase arrest in both HRSV-infected primary cells and A549 cells was confirmed using dual-label flow cytometry that accurately measured the different stages of the cell cycle. Comparison of progeny virus production in primary and A549 cells enriched in G(0)/G(1) using a specific CDK4/6 kinase inhibitor with asynchronously replicating cells indicated that this phase of the cell cycle was more efficient for virus production.
authors.affiliation: Institute of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Weining
authors.initials: W
authors.lastname: Wu
authors.forename: Diane C
authors.initials: DC
authors.lastname: Munday
authors.forename: Gareth
authors.initials: G
authors.lastname: Howell
authors.forename: Gareth
authors.initials: G
authors.lastname: Platt
authors.forename: John N
authors.initials: JN
authors.lastname: Barr
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000096:biosynthesis; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000821:virology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D006801:Humans; D018113:Respiratory Syncytial Virus, Human / Q000472:pathogenicity*; D015398:Signal Transduction
chemical_list: D018797:Cell Cycle Proteins