Phosphorylation of tick-borne encephalitis virus NS5 protein.
The largest tick-borne encephalitis virus (TBEV) non-structural protein NS5 (100 kDa) is believed to be involved in RNA replication. The protein is phosphorylated in infected cell extracts in the presence of [gamma-32P]ATP, as shown by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis using monoclonal antibodies raised against TBEV NS5 protein. Radioactive labeling of NS5 in cellular extracts at an early stage post-infection is higher than at 24 h post-infection. Incubation of immunoprecipitates of NS5 protein with [gamma-32P]ATP in the presence of Mg2+ resulted in the phosphorylation of TBEV NS5 protein and of immunoglobulins. Phosphoamino acid analysis demonstrated that NS5 contains phosphoserine, but not phosphothreonine, or phosphotyrosine.
Morozova
O V
OV
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Russia.
Tsekhanovskaya
N A
NA
Maksimova
T G
TG
Bachvalova
V N
VN
Matveeva
V A
VA
Kit YYa
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002478:Cells, Cultured; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004669:Encephalitis Viruses, Tick-Borne / Q000502:physiology*; D005260:Female; D007668:Kidney; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis; D010769:Phosphothreonine / Q000032:analysis; D019000:Phosphotyrosine / Q000032:analysis; D012367:RNA, Viral / Q000096:biosynthesis; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; C096041:NS5 protein, flavivirus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D010769:Phosphothreonine; D010768:Phosphoserine; D019000:Phosphotyrosine; D000255:Adenosine Triphosphate; D012321:DNA-Directed RNA Polymerases