Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Identification of Nuclear Factors that UV-crosslink to Rev-responsive Element RNA

UV조사에 의해 Rev-responsive element RNA와 결합하는 핵단백질인자의 확인

  • 박희성 (대구효성카톨릭대학교 식물육종학과) ;
  • 남용석 (고려대학교 의과대학 법의학교실)
  • Published : 1997.09.01
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Abstract

HIV-1 Rev protein plays an important role in regulating the expression of viral structural proteins. It allows the nuclear export and accumulation of unspliced and partially spliced viral mRNA in the cytoplasm. The Rev-responsive element RNA, present in the env gene, forms a higly ordered RNA secondary structure and is required for the Rev-mediated mRNA export. For this process to complete factor(s) are strongly suggested. From our experiments of electrophoretic mobility shift, UV-crosslinking and SDS/PAGE, RRE RNA was found to be recognized to several nuclear factors such as 36/37, 56, 41. 76, 150 kD proteins in the order of reactivity. Among them, 36/37 and 56 kD proteins are more reactive upon a brief UV treatment (5 min) and more persistent in the presence of high amount of nonspecific competitor, heparin. Certain nuclear protein9s) seemed to recognize the RRE RNA structure in competition with Rev to gel mobility shift assay.

HIV-1 Rev단백질은 바이러스의 구조단백질의 발현조절에 매우 중요한 역할을 하며RNA가 1차발현된 원형대로 또는 일부만이 절단/재조합된 상태로 세포질에 집적되는 것을 가능하게 한다. env gene에 존재하는 Rev-responsive element RNA는 복잡한 RNA구조를 지니면서 Rev의 기능을 위하여 필수적으로 필요시 된다. 그러나 이러한 절차의 완전한 진행을 위해서는 핵단백질이 요구되는 것으로 추정된다. 본 연구에서는 electrophoretic mobility shift, UV-crosslinking 또는 SDS/PAGE등의 실험을 통하여 36/37, 56, 41, 76, 150 kD등의 핵단백질등이 RRE RNA와 결합반응하는 것을 발견하였으며 특히 36/37 과 56 kD 단백질을 5분간의 자외선조사에 의해 특히 RRE RNA에 결합하는 핵단백질들은 gel mobility shift assay에서 Rev RNA인식결합에 경쟁적인 특징을 나타내고 있다.

Keywords

References

  1. Cell v.46 HTLV-Ⅲ experession and production involve complex regulation at the levels of splicing and translation of viral RNA. Feinberg, M. B.;Jarrett, R. F.;Aldovini, A.;Gallo, R. C.;Wong-Staal, F.
  2. Cell v.60 HIV-1 regulator of viron expression (Rev) binds to and RNA stem-loop structure located in the Rev responsive element region. Heapy, S.;Dingwall, C.;Ernberg, I.;Gait, M. J.;Green, S. M.;Karn, J.;Lowe, A. D.;Singh, M.;Skinner, M. A.
  3. Cell v.57 The recgene product of the HIV affects envelope-specific RNA localization. Emerman, M;Vazeur, R.;Penden, K.
  4. J. Virol. v.64 Human immunodificency virus rev protein recognizes a target sequence in revresponsive element RNA within the context of RNA econdary structure. Holland, S. M.;Ahmad, N.;Maitra, R. K.;Wingfield, P.;Venkatesan, S.
  5. Nature v.338 The HIV-1 Rev trans-activator : derivation of a trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA. Malim, M. H.;Hauber, J.;Le, S.-Y.;Maizel, J. V.;Cullen, B. R.
  6. Cell v.58 Functional dissection of the HIV-1 Rev transactivator-derivation of a trans-dominant repressor of Rev function. Malim, M. H.;Bohnlein, S.;Hauber, J.;Cullen, B. R.
  7. Cell v.57 A binding consensus : RNA-protein interactions in splicing, snRNPs, and sex. Mattaj, I. W.
  8. Cell v.59 Regulation by Rev dependes upon recognition of splice sites. Chang, D. D.;Sharp, P. A.
  9. Proc. Natl. Acad. Sci. USA v.87 Specific-interaction of the HIV Rev protein with a structured region in the env mRNA. Cochrane, A. W.;Chen, C.-H.;Rosen, C. A.
  10. Science v.247 Secondary structure is the major determinant for interaction of HIV rev protein with RNA. Olsen, H. S.;Nelbrook, P.;Cocharane, A. W.;Rosen, C. A.
  11. Proc. Natl. Acad. Sci. USA v.86 Rev protein human immunodeficiency virus type-1 affects the stability and transport of the viral mRNA. Felber, B. K.;Hadzopoulou-Cladaras, M.;Cladaras, C.;Copeland, T.;Pavlakis, G. N.
  12. Nature v.342 sequence-specific RNA binding by the HIV-1 Rev protein. Zapp, M. L.;Green, M. R.
  13. Science v.246 Functional analysis of CAR, the sequence for the Rev Protein of HIV-1. Dayton, E. T.;Powell, D. M.;Dayton, A. I.
  14. J. Virol. v.63 Subcellular localization of the HIV trans-acting art gene product. Cullen, B. R.;Hauber, J.;Campbell, K.;Sodroski, J. G.;Haseltine, W. A.;Rosen, C. A.
  15. Nucl. Acids Res. v.18 Identification of trans-dominant HIV-1 Rev protein mutants by direct transfer of bacterially produced proteins into human cells. Mermer, B.;Felber, B. K.;Campbell, M.;Pavalakis, G. N.
  16. J. Virol. v.65 Cellular factors regulate transactivation of HIV-1. Barry, P.;Pratt-Lowe, E.;Unger, R. E.;Luciw, P. A.
  17. New Biol. v.3 Identification and characterization of a nuclear factor that specifically binds to the Rev response element (RRE) of human immunodeficiency virus type-1 (HIV-1). Vaishnav, Y. N.;Vaishnav, M.;Wong-Staal, F.
  18. EMBO J. v.9 A human cell factor is essential for HIV-1 Rev protein. Trono, D.;Baltimore, D.
  19. Mol. Cell. Biol. v.11 Specific complex of human immunodeficiency virus type 1 Rev and nucleolar B 23 proteins : Dissociation by the Rev responsive element. Fankhauser, C.;Izaurralde, E.;Adachi, Y.;Wingfield, P.;Laemmli, U. K.
  20. Nucl. Acids Res. v.11 Accurate transcription initiation by RNA polymerase Ⅱ in a soluble extract from mammalian nuclei. Dingnam, J. D.;Lebowitz, R. M.;Roeder, R. G.
  21. Kor. J. Life Sci. v.6 RNA binding specificities of double-stranded RNA binding protein (RBF) as an inhibitor of PKR kinase. Park, H.;Choi, J.
  22. Proc. Natl. Acad. Sci. USA v.91 TAR RNA-binding protein is an inhibitor of the interferon-in-duced 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.
  23. Kor. J. Gen. v.19 Inhibition of PKR phosphorylation in vitro by Lac-Z fuxed double-stranded RNA binding protein (RBF) produced form E. coli. Park, H.;Choi, J.