[Phosphorylation of proteins in cytoplasmic and nuclear fractions of human cells treated with double-helical RNA and human recombinant interferon type I].
The dynamics of protein kinases activity in nuclear and cytoplasmic fractions of human fibroblasts treated by preparations of natural and synthetic dsRNA (ridostin, rifastin, larifan and poly(I).poly(C), DEAE-dextran and dsRNA complexes with DEAE-dextran), as well as by preparations of recombinant alpha-2 and beta-1 interferons was obtained. The early activation of enzymes in treated cells extracts and their presence in dsRNA-activated and nonactivated forms were found. In cytoplasmic cellular fractions treated by interferons the dsRNA dependent protein kinases (nonactivated forms- were prevalent.r In contrast, in dsRNA treated cells or dsRNA complexes with DEAE-dextran treated ones the dsRNA independent protein kinases (activated forms) were found, while dsRNA dependent forms induced by interferons were found at later periods. Nuclear protein kinases are mainly dsRNA independent making possible the supposition of their intracellular activation by incoming dsRNA or interferon-induced formation of ds-structures in cellular nuclei. In phosphorylated proteins spectre the 90, 69, 45-40 and 30-35 kDa polypeptides were found. At early intervals in nuclear fractions was found a nuclease resistant and partially EDTA resistant high molecular phosphorylated complex (120 kDa). The complex is, probably, capable of dissociation to low mol mass components. DEAE-dextran induces strong activation of protein kinases in cytoplasm and nuclei and increases the content of activated forms of enzyme in larifan treated cells.
Sokolova
T M
TM
Antonov
A Iu
AIu
D002467:Cell Nucleus / Q000201:enzymology; D003593:Cytoplasm / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D007369:Interferon Inducers / Q000494:pharmacology; D007370:Interferon Type I / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000378:metabolism*; D009930:Organic Chemicals; D010766:Phosphorylation; D011070:Poly I-C / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000378:metabolism*; D011994:Recombinant Proteins
D007369:Interferon Inducers; D007370:Interferon Type I; D009687:Nuclear Proteins; D009930:Organic Chemicals; D011506:Proteins; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; C066530:larifan; D011494:Protein Kinases; D011070:Poly I-C
Fosforilirovanie belkov v tsitoplazmaticheskoĭ i iadernoĭ fraktsiiakhchelovecheskikh kletok, obrabotannykh dvuspiral'nymi RNK i chelovecheskimi rekombinantnymi interferonami tipa 1.