Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Sep 10;11(17):5821–5835. doi: 10.1093/nar/11.17.5821

CAP binding proteins associated with the nucleus.

E Patzelt, D Blaas, E Kuechler
PMCID: PMC326320  PMID: 6889136

Abstract

Cap binding proteins of HeLa cells were identified by photo-affinity labelling using the cap analogue gamma-[32P]-[4-(benzoyl-phenyl)methylamido]-7-methylguanosine-5'- triphosphate. Photoreaction with whole cell homogenates resulted in specific labelling of five major polypeptides. The small molecular weight polypeptide appeared to be identical to the 24 000 to 26 000 dalton cap binding protein previously identified in initiation factors. A cap binding protein of 37 000 dalton was found in initiation factors as well as in preparations of crude nuclei. It was released from nuclei by washing with buffer of moderate salt concentration. Three high molecular weight cap binding proteins (approximately 120 000, approximately 89 000, approximately 80 000 dalton) were found in the nuclear fraction and were only partly released upon nuclease digestion and high salt extraction.

Full text

PDF
5823

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ben-Ze'ev A., Abulafia R., Aloni Y. SV40 virions and viral RNA metabolism are associated with cellular substructures. EMBO J. 1982;1(10):1225–1231. doi: 10.1002/j.1460-2075.1982.tb00017.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blaas D., Patzelt E., Kuechler E. Cap-recognizing protein of influenza virus. Virology. 1982 Jan 15;116(1):339–348. doi: 10.1016/0042-6822(82)90425-1. [DOI] [PubMed] [Google Scholar]
  3. Blaas D., Patzelt E., Kuechler E. Identification of the cap binding protein of influenza virus. Nucleic Acids Res. 1982 Aug 11;10(15):4803–4812. doi: 10.1093/nar/10.15.4803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown B. A., Ehrenfeld E. Translation of poliovirus RNA in vitro: changes in cleavage pattern and initiation sites by ribosomal salt wash. Virology. 1979 Sep;97(2):396–405. doi: 10.1016/0042-6822(79)90350-7. [DOI] [PubMed] [Google Scholar]
  5. Capco D. G., Wan K. M., Penman S. The nuclear matrix: three-dimensional architecture and protein composition. Cell. 1982 Jul;29(3):847–858. doi: 10.1016/0092-8674(82)90446-9. [DOI] [PubMed] [Google Scholar]
  6. Ciejek E. M., Nordstrom J. L., Tsai M. J., O'Malley B. W. Ribonucleic acid precursors are associated with the chick oviduct nuclear matrix. Biochemistry. 1982 Sep 28;21(20):4945–4953. doi: 10.1021/bi00263a018. [DOI] [PubMed] [Google Scholar]
  7. Etchison D., Milburn S. C., Edery I., Sonenberg N., Hershey J. W. Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220,000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. J Biol Chem. 1982 Dec 25;257(24):14806–14810. [PubMed] [Google Scholar]
  8. Grifo J. A., Tahara S. M., Leis J. P., Morgan M. A., Shatkin A. J., Merrick W. C. Characterization of eukaryotic initiation factor 4A, a protein involved in ATP-dependent binding of globin mRNA. J Biol Chem. 1982 May 10;257(9):5246–5252. [PubMed] [Google Scholar]
  9. Hansen J. L., Etchison D. O., Hershey J. W., Ehrenfeld E. Localization of cap-binding protein in subcellular fractions of HeLa cells. Mol Cell Biol. 1982 Dec;2(12):1639–1643. doi: 10.1128/mcb.2.12.1639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hansen J., Ehrenfeld E. Presence of the cap-binding protein in initiation factor preparations from poliovirus-infected HeLa cells. J Virol. 1981 May;38(2):438–445. doi: 10.1128/jvi.38.2.438-445.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hansen J., Etchison D., Hershey J. W., Ehrenfeld E. Association of cap-binding protein with eucaryotic initiation factor 3 in initiation factor preparations from uninfected and poliovirus-infected HeLa cells. J Virol. 1982 Apr;42(1):200–207. doi: 10.1128/jvi.42.1.200-207.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Herlan G., Eckert W. A., Kaffenberger W., Wunderlich F. Isolation and characterization of an RNA-containing nuclear matrix from Tetrahymena macronuclei. Biochemistry. 1979 May 1;18(9):1782–1788. doi: 10.1021/bi00576a023. [DOI] [PubMed] [Google Scholar]
  13. Herman R., Weymouth L., Penman S. Heterogeneous nuclear RNA-protein fibers in chromatin-depleted nuclei. J Cell Biol. 1978 Sep;78(3):663–674. doi: 10.1083/jcb.78.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jackson D. A., McCready S. J., Cook P. R. RNA is synthesized at the nuclear cage. Nature. 1981 Aug 6;292(5823):552–555. doi: 10.1038/292552a0. [DOI] [PubMed] [Google Scholar]
  15. Kuechler E., Barta A. Aromatic ketone derivatives of aminoacyl-tRNA as photoaffinity labels for ribosomes. Methods Enzymol. 1977;46:676–683. doi: 10.1016/s0076-6879(77)46085-3. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lebkowski J. S., Laemmli U. K. Non-histone proteins and long-range organization of HeLa interphase DNA. J Mol Biol. 1982 Apr 5;156(2):325–344. doi: 10.1016/0022-2836(82)90332-1. [DOI] [PubMed] [Google Scholar]
  18. Lee K. A., Sonenberg N. Inactivation of cap-binding proteins accompanies the shut-off of host protein synthesis by poliovirus. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3447–3451. doi: 10.1073/pnas.79.11.3447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Long B. H., Huang C. Y., Pogo A. O. Isolation and characterization of the nuclear matrix in Friend erythroleukemia cells: chromatin and hnRNA interactions with the nuclear matrix. Cell. 1979 Dec;18(4):1079–1090. doi: 10.1016/0092-8674(79)90221-6. [DOI] [PubMed] [Google Scholar]
  20. Mariman E. C., van Eekelen C. A., Reinders R. J., Berns A. J., van Venrooij W. J. Adenoviral heterogeneous nuclear RNA is associated with the host nuclear matrix during splicing. J Mol Biol. 1982 Jan 5;154(1):103–119. doi: 10.1016/0022-2836(82)90420-x. [DOI] [PubMed] [Google Scholar]
  21. Mariman E., Hagebols A. M., van Venrooij W. On the localization and transport of specific adenoviral mRNA-sequences in the late infected HeLa cell. Nucleic Acids Res. 1982 Oct 11;10(19):6131–6145. doi: 10.1093/nar/10.19.6131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maundrell K., Maxwell E. S., Puvion E., Scherrer K. The nuclear matrix of duck erythroblasts is associated with globin mRNA coding sequences but not with the major proteins of 40S nuclear RNP. Exp Cell Res. 1981 Dec;136(2):435–445. doi: 10.1016/0014-4827(81)90023-9. [DOI] [PubMed] [Google Scholar]
  23. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  24. Plotch S. J., Bouloy M., Ulmanen I., Krug R. M. A unique cap(m7GpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription. Cell. 1981 Mar;23(3):847–858. doi: 10.1016/0092-8674(81)90449-9. [DOI] [PubMed] [Google Scholar]
  25. Ray B. K., Brendler T. G., Adya S., Daniels-McQueen S., Miller J. K., Hershey J. W., Grifo J. A., Merrick W. C., Thach R. E. Role of mRNA competition in regulating translation: further characterization of mRNA discriminatory initiation factors. Proc Natl Acad Sci U S A. 1983 Feb;80(3):663–667. doi: 10.1073/pnas.80.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ross D. A., Yen R. W., Chae C. B. Association of globin ribonucleic acid and its precursors with the chicken erythroblast nuclear matrix. Biochemistry. 1982 Feb 16;21(4):764–771. doi: 10.1021/bi00533a029. [DOI] [PubMed] [Google Scholar]
  27. Schreier M. H., Staehelin T. Initiation of mammalian protein synthesis: the importance of ribosome and initiation factor quality for the efficiency of in vitro systems. J Mol Biol. 1973 Feb 19;73(3):329–349. doi: 10.1016/0022-2836(73)90346-x. [DOI] [PubMed] [Google Scholar]
  28. Sonenberg N. ATP/Mg++-dependent cross-linking of cap binding proteins to the 5' end of eukaryotic mRNA. Nucleic Acids Res. 1981 Apr 10;9(7):1643–1656. doi: 10.1093/nar/9.7.1643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sonenberg N., Guertin D., Cleveland D., Trachsel H. Probing the function of the eucaryotic 5' cap structure by using a monoclonal antibody directed against cap-binding proteins. Cell. 1981 Dec;27(3 Pt 2):563–572. doi: 10.1016/0092-8674(81)90398-6. [DOI] [PubMed] [Google Scholar]
  30. Sonenberg N., Morgan M. A., Merrick W. C., Shatkin A. J. A polypeptide in eukaryotic initiation factors that crosslinks specifically to the 5'-terminal cap in mRNA. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4843–4847. doi: 10.1073/pnas.75.10.4843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sonenberg N., Rupprecht K. M., Hecht S. M., Shatkin A. J. Eukaryotic mRNA cap binding protein: purification by affinity chromatography on sepharose-coupled m7GDP. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4345–4349. doi: 10.1073/pnas.76.9.4345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sonenberg N., Trachsel H. Probing the function of the eukaryotic 5'-cap structure using monoclonal antibodies to cap-binding proteins. Curr Top Cell Regul. 1982;21:65–88. doi: 10.1016/b978-0-12-152821-8.50007-4. [DOI] [PubMed] [Google Scholar]
  33. Tahara S. M., Morgan M. A., Shatkin A. J. Two forms of purified m7G-cap binding protein with different effects on capped mRNA translation in extracts of uninfected and poliovirus-infected HeLa cells. J Biol Chem. 1981 Aug 10;256(15):7691–7694. [PubMed] [Google Scholar]
  34. Ulmanen I., Broni B. A., Krug R. M. Role of two of the influenza virus core P proteins in recognizing cap 1 structures (m7GpppNm) on RNAs and in initiating viral RNA transcription. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7355–7359. doi: 10.1073/pnas.78.12.7355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zumbé A., Stähli C., Trachsel H. Association of a Mr 50,000 cap-binding protein with the cytoskeleton in baby hamster kidney cells. Proc Natl Acad Sci U S A. 1982 May;79(9):2927–2931. doi: 10.1073/pnas.79.9.2927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. van Eekelen C. A., van Venrooij W. J. hnRNA and its attachment to a nuclear protein matrix. J Cell Biol. 1981 Mar;88(3):554–563. doi: 10.1083/jcb.88.3.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. van Eekelen C., Ohlsson R., Philipson L., Mariman E., van Beek R., van Venrooij W. Sequence dependent interaction of hnRNP proteins with late adenoviral transcripts. Nucleic Acids Res. 1982 Nov 25;10(22):7115–7131. doi: 10.1093/nar/10.22.7115. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES