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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1977 Apr;74(4):1426–1430. doi: 10.1073/pnas.74.4.1426

Heterogeneous nuclear RNA secondary structure: oligo (U) sequences base-paired with poly (A) and their possible role as binding sites for heterogeneous nuclear RNA-specific proteins.

V M Kish, T Pederson
PMCID: PMC430784  PMID: 266183

Abstract

HeLa cell heterogeneous nuclear RNA derived from high-molecular-weight nuclear ribonucleoprotein (RNP) particles contains oligo(U) sequences of 15-50 nucleotides base-paired with poly(A). These duplexes are resistant to pancreatic RNase at 0.5 M NaCl in native RNP, remain so after chemical deproteinization of the RNP digests, and then copurify with poly(A) on oligo(dT)-cellulose chromatography. Oligo(dT)-cellulose binding capacity of the oligo(U)-poly(A) duplexes is abolished by prior titration of the nonduplex poly(A) regions with excess poly(U). The oligo(dT)-purified fraction is 97.5 mole % A + U and the [3H]uridine-labeled component is resistant to redigestion by pancreatic RNase at 0.5 M NaCl but not at 0.01 M NaCl. After thermal denaturation, the [3H]uridine-labeled chains become RNase-sensitive at 0.5 M NaCl. Electrophoresis of [3H]adenosine- or [3H]uridine-labeled material in polyacrylamide gels containing 99% formamide confirms that the oligo(U) sequences are not covalently linked to poly(A). Controls establish that the A-U duplexes are not formed artifactually during isolation of heterogeneous nuclear RNP or subsequent fractionation. The oligo(U)-poly(A) duplexes appear to be associated with protein in native heterogeneous nuclear RNP, as reflected by the differential pancreatic RNase sensitivity of the duplexed oligo(U) in RNP (resistant) and RNA (sensitive), measured at physiological ionic strength.

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Selected References

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

  1. BEERS R. F., Jr Hydrolysis of polyadenylic acid by pancreatic ribonuclease. J Biol Chem. 1960 Aug;235:2393–2398. [PubMed] [Google Scholar]
  2. Darnell J. E., Philipson L., Wall R., Adesnik M. Polyadenylic acid sequences: role in conversion of nuclear RNA into messenger RNA. Science. 1971 Oct 29;174(4008):507–510. doi: 10.1126/science.174.4008.507. [DOI] [PubMed] [Google Scholar]
  3. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Edy V. G., Szekely M., Loviny T., Dreyer C. Action of nucleases on double-stranded RNA. Eur J Biochem. 1976 Jan 15;61(2):563–572. doi: 10.1111/j.1432-1033.1976.tb10051.x. [DOI] [PubMed] [Google Scholar]
  5. Firtel R. A., Pederson T. Ribonucleoprotein particles containing heterogeneous nuclear RNA in the cellular slime mold Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 Jan;72(1):301–305. doi: 10.1073/pnas.72.1.301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jelinek W., Darnell J. E. Double-stranded regions in heterogeneous nuclear RNA from Hela cells. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2537–2541. doi: 10.1073/pnas.69.9.2537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kish V. M., Pederson T. Poly (A)-rich ribonucleoprotein complexes from HeLa cell messenger RNA. J Biol Chem. 1976 Oct 10;251(19):5888–5894. [PubMed] [Google Scholar]
  8. Kish V. M., Pederson T. Ribonucleoprotein organization of polyadenylate sequences in HeLa cell heterogeneous nuclear RNA. J Mol Biol. 1975 Jun 25;95(2):227–238. doi: 10.1016/0022-2836(75)90392-7. [DOI] [PubMed] [Google Scholar]
  9. Korwek E. L., Nakazato N., Edmonds M., Venkatesan S. Poly(uridylic acid) sequences in messenger ribonucleic acid of HeLa cells. Biochemistry. 1976 Oct 19;15(21):4643–4649. doi: 10.1021/bi00666a015. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Mendecki J., Lee S. Y., Brawerman G. Characteristics of the polyadenylic acid segment associated with messenger ribonucleic acid in mouse sarcoma 180 ascites cells. Biochemistry. 1972 Feb 29;11(5):792–798. doi: 10.1021/bi00755a018. [DOI] [PubMed] [Google Scholar]
  12. Molloy G. R., Darnell J. E. Characterization of the poly(adenylic acid) regions and the adjacent nucleotides in heterogeneous nuclear ribonucleic acid and messenger ribonucleic acid from HeLa cells. Biochemistry. 1973 Jun 5;12(12):2324–2330. doi: 10.1021/bi00736a022. [DOI] [PubMed] [Google Scholar]
  13. Molloy G. R., Thomas W. L., Darnell J. E. Occurrence of uridylate-rich oligonucleotide regions in heterogeneous nuclear RNA of HeLa cells. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3684–3688. doi: 10.1073/pnas.69.12.3684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakazato H., Edmonds M., Kopp D. W. Differential metabolism of large and small poly(A) sequences in the heterogeneous nuclear RNA of HeLa cells. Proc Natl Acad Sci U S A. 1974 Jan;71(1):200–204. doi: 10.1073/pnas.71.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakazato H., Kopp D. W., Edmonds M. Localization of the polyadenylate sequences in messenger ribonucleic acid and in the heterogeneous nuclear ribonucleic acid of HeLa cells. J Biol Chem. 1973 Feb 25;248(4):1472–1476. [PubMed] [Google Scholar]
  16. Pederson T. Proteins associated with heterogeneous nuclear RNA in eukaryotic cells. J Mol Biol. 1974 Feb 25;83(2):163–183. doi: 10.1016/0022-2836(74)90386-6. [DOI] [PubMed] [Google Scholar]
  17. Riley M., Maling B. Physical and chemical characterization of two- and three-stranded adenine-thymine and adenine-uracil homopolymer complexes. J Mol Biol. 1966 Sep;20(2):359–389. doi: 10.1016/0022-2836(66)90069-6. [DOI] [PubMed] [Google Scholar]
  18. Ryskov A. P., Saunders G. F., Farashyan V. R., Georgiev G. P. Double-helical regions in nuclear precursor of mRNA (pre-mRNA). Biochim Biophys Acta. 1973 Jun 8;312(1):152–164. doi: 10.1016/0005-2787(73)90060-9. [DOI] [PubMed] [Google Scholar]

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