Vimentin rearrangement during African swine fever virus infection involves retrograde transport along microtubules and phosphorylation of vimentin by calcium calmodulin kinase II.
African swine fever virus (ASFV) infection leads to rearrangement of vimentin into a cage surrounding virus factories. Vimentin rearrangement in cells generally involves phosphorylation of N-terminal domains of vimentin by cellular kinases to facilitate disassembly and transport of vimentin filaments on microtubules. Here, we demonstrate that the first stage in vimentin rearrangement during ASFV infection involves a microtubule-dependent concentration of vimentin into an "aster" within virus assembly sites located close to the microtubule organizing center. The aster may play a structural role early during the formation of the factory. Conversion of the aster into a cage required ASFV DNA replication. Interestingly, viral DNA replication also resulted in the activation of calcium calmodulin-dependent protein kinase II (CaM kinase II) and phosphorylation of the N-terminal domain of vimentin on serine 82. Immunostaining showed that vimentin within the cage was phosphorylated on serine 82. Significantly, both viral DNA replication and Ser 82 phosphorylation were blocked by KN93, an inhibitor of CaM kinase II, suggesting a link between CaM kinase II activation, DNA replication, and late gene expression. Phosphorylation of vimentin on serine 82 may be necessary for cage formation or may simply be a consequence of activation of CaM kinase II by ASFV. The vimentin cage may serve a cytoprotective function and prevent movement of viral components into the cytoplasm and at the same time concentrate late structural proteins at sites of virus assembly.
Stefanovic
Sandra
S
Division of Immunology, Pirbright Laboratories, Institute for Animal Health, Surrey Ash Road, Woking, Surrey GU24 ONF, United Kingdom.
Windsor
Miriam
M
Nagata
Koh-Ici
KI
Inagaki
Masaki
M
Wileman
Thomas
T
D000357:African Swine Fever / Q000378:metabolism* / Q000821:virology*; D000358:African Swine Fever Virus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D000818:Animals; D001693:Biological Transport, Active; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D004789:Enzyme Activation; D015870:Gene Expression; D005814:Genes, Viral; D008870:Microtubules / Q000378:metabolism; D010766:Phosphorylation; D012694:Serine / Q000737:chemistry; D013552:Swine; D014709:Vero Cells; D014746:Vimentin / Q000737:chemistry / Q000378:metabolism*; D019065:Virus Assembly
D004279:DNA, Viral; D014746:Vimentin; D012694:Serine; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases