TICK-BORNE ENCEPHALITIS VIRUS REPLICATION COMPLEX. I. IDENTIFICATION OF A PROTEIN RESPONSIBLE FOR THE INITIATION OF RNA SYNTHESIS IN NUCLEAR-ASSOCIATED FRACTION OF INFECTED CELLS BY MEANS OF AFFINITY LABELLING
O. V. MOROZOVA, N. A. BELYAVSKAYA, E. F. ZAYCHIKOV, E. A. KVETKOVA, A. A. MUTAEV, A. G. PLETNEV
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Academy of Sciences of the USSR; Omsk Institute of Natural-Foci Infections, Ministry of Health of the RSFSR; Immnological Institute, Siberian Division of the Academy of Sciences of the USSR, Irkutsk
Abstract: The pig embrio kidney cells infected by tick-borne encephalitis virus were fractionated into nuclear-associated, cytoplasmic and membrane fractions. The main part of the virus replicase activity was associated with the nuclei. The replication complex is able to synthesize full-length viral RNAs in vitro. To identify proteins involved in the initiation of the replication at the late stages of the infection, the highly specific affinity labelling technique was used. It was shown that with aldehyde-containing derivatives of ATP, ADP and AMP and [α-32P]GTP the target of labelling is a polypeptide having molecular weight about 69 kDa. The same protein is immunostained with TBE virus specific antibodies after blotting onto nitrocellulose. The conclusion is made that nonstructural protein NS3 takes part in virus replication at the late-stage of the infection.
Russian Journal of Bioorganic Chemistry 1990, 16 (4):492-500