Protein kinase R is responsible for the phosphorylation of eIF2alpha in rotavirus infection.
The eukaryotic initiation translation factor 2 (eIF2) represents a key point in the regulation of protein synthesis. This factor delivers the initiator Met-tRNA to the ribosome, a process that is conserved in all eukaryotic cells. Many types of stress reduce global translation by triggering the phosphorylation of the α subunit of eIF2, which reduces the formation of the preinitiation translation complexes. Early during rotavirus infection, eIF2α becomes phosphorylated, and even under these conditions viral protein synthesis is not affected, while most of the cell protein synthesis is blocked. Here, we found that the kinase responsible for the phosphorylation of eIF2α in rotavirus-infected cells is PKR, since in mouse embryonic fibroblasts deficient in the kinase domain of PKR, or in MA104 cells where the expression of PKR was knocked down by RNA interference, eIF2α was not phosphorylated upon rotavirus infection. The viral component responsible for the activation of PKR seems to be viral double-stranded RNA, which is found in the cytoplasm of infected cells, outside viroplasms. Taken together, these results suggest that rotaviruses induce the PKR branch of the interferon system and have evolved a mechanism to translate its proteins, surpassing the block imposed by eIF2α phosphorylation.
Rojas
Margarito
M
Departamento de Génetica del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, UNAM, Cuernavaca, Morelos 62210, México.
Arias
Carlos F
CF
López
Susana
S
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D054884:Host-Pathogen Interactions / Q000502:physiology; D008253:Macaca mulatta; D051379:Mice; D018345:Mice, Knockout; D050505:Mutant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism / Q000821:virology; D012401:Rotavirus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology; D012400:Rotavirus Infections / Q000378:metabolism*; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis; D019892:eIF-2 Kinase / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
D015852:Eukaryotic Initiation Factor-2; D050505:Mutant Proteins; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D014764:Viral Proteins; D019892:eIF-2 Kinase