ABSTRACT
We studied influenza virus M1 protein by generating HeLa and MDCK cell lines that express M1 genetically fused to green fluorescent protein (GFP). GFP-M1 was incorporated into virions produced by influenza virus infected MDCK cells expressing the fusion protein indicating that the fusion protein is at least partially functional. Following infection of either HeLa or MDCK cells with influenza A virus (but not influenza B virus), GFP-M1 redistributes from its cytosolic/nuclear location and accumulates in nuclear dots. Immunofluorescence revealed that the nuclear dots represent nuclear dot 10 (ND10) structures. The colocalization of authentic M1, as well as NS1 and NS2 protein, with ND10 was confirmed by immunofluorescence following in situ isolation of ND10. These findings demonstrate a previously unappreciated involvement of influenza virus with ND10, a structure involved in cellular responses to immune cytokines as well as the replication of a rapidly increasing list of viruses.
Subject(s)
Cell Nucleus/chemistry , Orthomyxoviridae/metabolism , Viral Matrix Proteins/metabolism , Animals , Dogs , Green Fluorescent Proteins , HeLa Cells , Humans , Luminescent Proteins/analysis , Viral Matrix Proteins/analysis , Viral Nonstructural Proteins/analysis , Virion/metabolismABSTRACT
A new rotary-shadowing process to obtain freeze-drying replicas is described for the analysis of virus ultrastructure, using the inner capsid of human rotavirus as a model. The findings corroborate the icosahedral symmetry with an arrangement pattern of capsomers of T=13L
