Dynamics and interactions of parvoviral NS1 protein in the nucleus.
Nuclear positioning and dynamic interactions of viral proteins with nuclear substructures play essential roles during infection with DNA viruses. Visualization of the intranuclear interactions and motility of the parvovirus replication protein (NS1) in living cells gives insight into specific parvovirus protein-cellular structure interactions. Confocal analysis of highly synchronized infected Norden Laboratory Feline Kidney cells showed accumulation of nuclear NS1 in discrete interchromosomal foci. NS1 fused with enhanced yellow fluorescence protein (NS1-EYFP) provided a marker in live cells for dynamics of NS1 traced by photobleaching techniques. Fluorescence Recovery after Photobleaching suggested that the NS1 protein is not freely diffusing but undergoes transient interactions with nuclear compartments. Fluorescence Loss in Photobleaching demonstrated for the first time the shuttling of a parvoviral protein between the nucleus and the cytoplasm as assayed with NS1-EYFP. Finally, time-lapse imaging of infected cells revealed that the intranuclear distribution of NS1-EYFP evolves dramatically starting from the formation of NS1 foci and proceeding to a homogenous distribution extending throughout the nucleus.
Ihalainen
Teemu O
TO
Department of Biological and Environmental Science, NanoScience Center, University of Jyväskylä, Survontie 9, FI-40014 Jyväskylä, Finland.
Niskanen
Einari A
EA
Jylhävä
Juulia
J
Turpeinen
Tuomas
T
Rinne
Johanna
J
Timonen
Jussi
J
Vihinen-Ranta
Maija
M
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002415:Cats; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D017992:Parvovirus / Q000378:metabolism / Q000502:physiology*; D038761:Photobleaching; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D017361:Viral Nonstructural Proteins; C054562:yellow fluorescent protein, Bacteria