Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1979 Sep 11;7(1):135–149. doi: 10.1093/nar/7.1.135

Purification of histone messenger ribonucleoprotein particles from HeLa cell S-phase polysomes. Characterization of associated proteins

JP Liautard 1, Ph Jeanteur 2
PMCID: PMC328001  PMID: 493137

Abstract

We have purified HeLa histone mRNA from polysomes of S-phase cells which had been synchronized by hydroxyurea treatment. This mRNA was shown to direct the in vitro synthesis of all five histones which amount to at least 90-95% of its total translational activity. Polysomal histone mRNP was also purified and identified by cell-free translation and hybridization to a clone of histone DNA from E. esculentus. The protein moiety of this mRNP contained three prominent species of molecular weight 86,000, 73,000 and 53,000 daltons. The presence of the 73,000 species previously assessed to be bound to poly(A) is discussed in view of the fact that histone mRNA does not contain a poly(A) tail. As globin mRNA, histone mRNA as well as histone mRNP were translated with equal efficiency in cell-free extracts from either S-phase or hydroxyurea blocked HeLa cells.

Full text

PDF
136

Images in this article

Selected References

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

  1. Adesnik M., Darnell J. E. Biogenesis and characterization of histone messenger RNA in HeLa cells. J Mol Biol. 1972 Jun 28;67(3):397–406. doi: 10.1016/0022-2836(72)90458-5. [DOI] [PubMed] [Google Scholar]
  2. Auerbach S., Pederson T. Phosphorylation of messenger RNA-bound proteins in HeLa cells. Biochem Biophys Res Commun. 1975 Mar 3;63(1):149–156. doi: 10.1016/s0006-291x(75)80023-4. [DOI] [PubMed] [Google Scholar]
  3. Blobel G. A protein of molecular weight 78,000 bound to the polyadenylate region of eukaryotic messenger RNAs. Proc Natl Acad Sci U S A. 1973 Mar;70(3):924–928. doi: 10.1073/pnas.70.3.924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blobel G. Protein tightly bound to globin mRNA. Biochem Biophys Res Commun. 1972 Apr 14;47(1):88–95. doi: 10.1016/s0006-291x(72)80014-7. [DOI] [PubMed] [Google Scholar]
  5. Bryan R. N., Hayashi M. Two proteins are bound to most species of polysomal mRNA. Nat New Biol. 1973 Aug 29;244(139):271–274. doi: 10.1038/newbio244271a0. [DOI] [PubMed] [Google Scholar]
  6. Chen J. H., Lavers G. C., Spector A. Calf lens messenger ribonucleoprotein complexes. Characterization and comparison of template activity with corresponding mRNAs. Biochim Biophys Acta. 1976 Jan 5;418(1):39–51. doi: 10.1016/0005-2787(76)90325-7. [DOI] [PubMed] [Google Scholar]
  7. Ducamp C., Jeanteur P. Characterization of nuclear RNP particles from Hela cells. Analysis of protein and RNA constituents. Presence of poly (A). Biochimie. 1973;55(10):1235–1243. doi: 10.1016/s0300-9084(74)80328-7. [DOI] [PubMed] [Google Scholar]
  8. Gedamu L., Dixon G. H., Davies P. L. Identification and isolation of protamine messenger ribonucleoprotein particles from rainbow trout testis. Biochemistry. 1977 Apr 5;16(7):1383–1391. doi: 10.1021/bi00626a023. [DOI] [PubMed] [Google Scholar]
  9. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  10. Greenberg J. R. Isolation of messenger ribonucleoproteins in cesium sulfate density gradients: evidence that polyadenylated and non-polyadenylated messenger RNAs are associated with protein. J Mol Biol. 1976 Dec;108(2):403–416. doi: 10.1016/s0022-2836(76)80127-1. [DOI] [PubMed] [Google Scholar]
  11. Greenberg J. R. Ultraviolet light-induced crosslinking of mRNA to proteins. Nucleic Acids Res. 1979 Feb;6(2):715–732. doi: 10.1093/nar/6.2.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Henshaw E. C. Messenger RNA in rat liver polyribosomes: evidence that it exists as ribonucleoprotein particles. J Mol Biol. 1968 Sep 28;36(3):401–411. doi: 10.1016/0022-2836(68)90164-2. [DOI] [PubMed] [Google Scholar]
  13. Irwin D., Kumar A., Malt R. A. Messenger ribonucleoprotein complexes isolated with oligo(dT)-cellulose chromatography from kidney polysomes. Cell. 1975 Feb;4(2):157–165. doi: 10.1016/0092-8674(75)90122-1. [DOI] [PubMed] [Google Scholar]
  14. Kumar A., Pederson T. Comparison of proteins bound to heterogeneous nuclear RNA and messenger RNA in HeLa cells. J Mol Biol. 1975 Aug 15;96(3):353–365. doi: 10.1016/0022-2836(75)90165-5. [DOI] [PubMed] [Google Scholar]
  15. Lebleu B., Marbaix G., Huez G., Temmerman J., Burny A., Chantrenne H. Characterization of the messenger ribonucleoprotein released from reticulocyte polyribosomes by EDTA treatment. Eur J Biochem. 1971 Mar 11;19(2):264–269. doi: 10.1111/j.1432-1033.1971.tb01313.x. [DOI] [PubMed] [Google Scholar]
  16. Liautard J. P., Setyono B., Spindler E., Köhler K. Comparison of proteins bound to the different functional classes of messenger RNA. Biochim Biophys Acta. 1976 Apr 2;425(4):373–383. doi: 10.1016/0005-2787(76)90001-0. [DOI] [PubMed] [Google Scholar]
  17. Liautard J. P., Tromm D., Köhler K. Isolation of messenger ribonucleoproteins from HeLa cells by affinity chromatography on poly (U) sepharose. Z Naturforsch C. 1977 May-Jun;32(5-6):419–423. doi: 10.1515/znc-1977-5-617. [DOI] [PubMed] [Google Scholar]
  18. Lindberg U., Sundquist B. Isolation of messenger ribonucleoproteins from mammalian cells. J Mol Biol. 1974 Jun 25;86(2):451–468. doi: 10.1016/0022-2836(74)90030-8. [DOI] [PubMed] [Google Scholar]
  19. Melli M., Spinelli G., Wyssling H., Arnold E. Presence of histone mRNA sequences in high molecular weight RNA of HeLa cells. Cell. 1977 Jul;11(3):651–661. doi: 10.1016/0092-8674(77)90082-4. [DOI] [PubMed] [Google Scholar]
  20. Milcarek C., Price R., Penman S. The metabolism of a poly(A) minus mRNA fraction in HeLa cells. Cell. 1974 Sep;3(1):1–10. doi: 10.1016/0092-8674(74)90030-0. [DOI] [PubMed] [Google Scholar]
  21. Morel Carlos, Kayibanda Boniface, Scherrer Klaus. Proteins associated with globin messenger RNA in avian erythroblasts: Isolation and comparison with the proteins bound to nuclear messenger-likie RNA. FEBS Lett. 1971 Oct 15;18(1):84–88. doi: 10.1016/0014-5793(71)80413-1. [DOI] [PubMed] [Google Scholar]
  22. Peacock A. C., Dingman C. W. Resolution of multiple ribonucleic acid species by polyacrylamide gel electrophoresis. Biochemistry. 1967 Jun;6(6):1818–1827. doi: 10.1021/bi00858a033. [DOI] [PubMed] [Google Scholar]
  23. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  24. Perry R. P., Kelley D. E. Messenger RNA-protein complexes and newly synthesized ribosomal subunits: analysis of free particles and components of polyribosomes. J Mol Biol. 1968 Jul 14;35(1):37–59. doi: 10.1016/s0022-2836(68)80035-x. [DOI] [PubMed] [Google Scholar]
  25. Stephens R. E., Pan C. J., Ajiro K., Dolby T. W., Borun T. W. Studies of human histone messenger RNA. I. Methods for the isolation and partial characterization of RNA fractions containing human histone message from HeLa S3 polyribosomes. J Biol Chem. 1977 Jan 10;252(1):166–172. [PubMed] [Google Scholar]
  26. Sundquist B., Persson T., Lindberg U. Characterization of mRNA-protein complexes from mammalian cells. Nucleic Acids Res. 1977 Apr;4(4):899–915. doi: 10.1093/nar/4.4.899. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES