RNA Binding Specificities of Double-Stranded RNA Binding Protein (RBF) as an Inhibitor of PRK Kinase

PKR인산화효소 억제인자인 이중선RNA결합단백질 (RBF)의 RNA결합특이성

  • 박희성 (대구효성카톨릭대학교 식물육종학과) ;
  • 최장원 (대구대학교 농화학과)
  • Published : 1996.12.01


A double-stranded RNA binding factor (RBF), characterized as an inhibitor of PKR kinase in our previous study, was evaluated for its RNA binding specificities by RNA gel electrophoretic mobility shift analysis and membrane filter binding assay, RBF displayed affinities for a broad range of RNAs including viral RNAs and synthetic RNAs consiting of stem and loop structures. GC-rich RNA stem helices as short as 11 bp are suggested to represent the minimal binding motif for RBF. RBF binding to all the natural RNAs tested was reversible by poly(I): poly(C) addition, but E. coli 5S RNA was inefficient.

PKR인산화효소의 억제인자로서 밝혀진 이중선RNA결합단백질 (RBF)의 RNA결합특이성을 정기영도에 의한 RNA 이동변화실험과 여과막결합도실험에 의해 측정하였다. RBF는 바이러스RNA나 stem/loop구조를 지니는 합성 RNA들에 대한 다양한 친화력을 지니는 것으로 나타났으며 충분한 GC가 포함된 11염기쌍으로 이루어진 RNA stem helix RBF가 결합하기 위한 최소한의 RNA구조로 제시되고 있다. 자연적 RNA구조에 대한 RBF의 결합은 poly(I) : poly(C)의 첨가에 의해 반전되었으며 E. coli 5S RNA경우는 효과를 거의 나타내지 않았다.


  1. Genes & Dev. v.3 RNA-binding proteins as developmental regulators Bandziulis, R. M.;Swanson, M. S.;Dreyfuss, G.
  2. Mol. Biol. Rep. v.14 A selective revies of RNA-protein interactions in eukaryotes Mattaj, I. W.
  3. Nucleic Acids Res. v.19 Detailed mutational analysis of TAR RNA : critical spacing between the bulge and loop recognition domains Berkout, B.;Jeang, K. T.
  4. Cell v.59 Sequence-specific recognition of RNA hairpins by bacteriophage antitermination requires a conseved arginine-rich motif Lazinski, D. E.;Grzadzielska, E.;Das, A.
  5. Mol. Cell. Biol. v.11 Molecular analysis of two poly(A) site-processing factors that determine the recognition and efficiency of cleavage of the pre-mRNA Gilmartin, G.;Nevins, J.
  6. Science v.240 Iron-responsive elements : regulatory RNA sequences that control mRNA levels and translation Casey, J. L.;Hentze, M. W.;Koeller, D. M.;Caughman, S. W.;Rouault, T. A.;Klausner, R. D.;Harford, J. B.
  7. Cell v.68 Phosphorylation of initiation factor 2 alpha by protein kinase GCN2 mediates gene-specific translational control of GCN4 in yeast Dever, T. E.;Feng, L.;Wek, R. C.;Cigan, A. M.;Donahue, T. F.;Hinnenbusch, A. G.
  8. Cell v.59 Tat trans-activates the human immunodeficiency virus through a nascent RNA target Berkout, B.;Silverman, R. H.;Jeang, K. T.
  9. J. Interferon Res. v.11 Interferon-induced and double-stranded RNA-activated enzymes : a specific protein kinase and 2',5'-oligoadenylate synthetases Hovanessian, A. G.
  10. Proc. Natl. Acad. Sci. USA v.76 Mechanism of interferon action : phosphorylation of protein synthesis initiation factor eIF-2 in interferon-treated human cells by a ribosome-associated kinase processing site specificity similar to hemin-regulated rabbit reticulocyte kinase Samuel, C. E.
  11. Annu. Rev. Biochem. v.56 Impact of virus infection on host cell protein synthesis Schneider, R. J.;Shenk, T.
  12. Eur. J. Biochem. v.178 The binding of double-stranded RNA and adenovirus VAI RNA to the interferon-induced protein kinase Galabru, J.;Katze, M. G.;Robert, N.;Hovanessian, A. G.
  13. Annu. Rev. Biochem. v.60 Translational control in mammalian cells Hershey, J. W. B.
  14. Virology v.172 Mechanism of interferon action. Activation of the human P1/eIF-2 alpha protein kinase by individual reovirus s-class mRNAs : s1 mRNA is a potent activator relative to s4 mRNA Bishoff, J.;Samuel, C. E.
  15. EMBO J. v.11 Human p68 kinase exhibits growth suppression in yeast and homology to the translational regulator GCN2 Chong, K. L.;Feng, L.;Schappert, K.;Meurs, E.;Donahue, T. F.;Friesen, J. D.;Hovanessian, A. G.;Williams, B. R.
  16. J. Virol. v.66 The Vaccinia virus K3L gene product potentiates translation by inhibiting dsRNA-activated protein kinase and phosphorylation of the alpha subunit of eukaryotic initiation factor 2 Davies, M. V.;Elroy-Stein, O.;Jagus, R.;Moss, B.;Kaufman, R.
  17. Mol. Cell. Biol. v.11 Partial characterization of a cellular factor that regulates the dsRNA-dependent eIF-2α kinase in 3T3-F442A fibroblasts Judware, R.;Petryshyn, R.
  18. J. Biol. Chem. v.267 Characterization and regulation of the 58, OOO-dalton cellular inhibitor of the interferon-induced, dsRNA-activated protein kiasne Lee, T. G.;Yomita, J.;Hovanessian, A. G.;Katze, M. G.
  19. Proc. Natl. Acad. Sci. USA v.91 TAR RNA-binding protein is an inhibitor of the interferon-induced protein kinase PKR Park, H.;Davies, M. V.;Langland, J. O.;Chang, H. W.;Nam, Y. S.;Tartaglia, J.;Paoletti, E.;Jacobs, B. L.;Kaufman, R. J.;Venkatesan, S.
  20. Kor. J. Gen. Inhibition of PKR phosphorylation in vitro by lacZ-fused double-stranded RNA binding protein (RBF) produced from E. coli Park, H.;Choi, J.
  21. J. Virol. v.64 Human immunodeficiency virus rev protein recognizes a target sequence in RRE RNA within the context of RNA secondary structure Holland, S. M.;Ahmad, N.;Maitra, R. K.;Wingfield, P.;Venkatesan, S.
  22. J. Virol. v.65 Adenovirus virus-associated RNA and translation control Matthews, M. B.;Shenk, T.
  23. Cell v.64 The HTLV-1 Rex responsive element mediates a novel form of mRNA polyadenylation Ahmad, Y. F.;Gilmartin, G. M.;Hanley, S. M.;Nevins, J. R.;Greene, W. C.
  24. J. Virol. v.66 Human immunodeficiency virus type 1 Rev activation can be achieved without RRE RNA if Rev is directed to the target as a Rev/MS2 fusion protein which tethers the MS2 operator RNS Venkatesan, S.;Gerstberger, S. M.;Park, H.;Holland, S. M.;Nam, Y.
  25. Proc. Natl. Acad. Sci. USA v.89 Identification of double stranded RNA binding domains in the interferon induced double stranded RNA activated p68 kinase Feng, G. S.;Chong, K.;Kumar, A.;Williams, B. R.
  26. Genes & Dev. v.6 Two RNA binding motifs in the double-stranded RNA activated protein kinase, DAI Green, S. R.;Mathews, M. B.