Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1980 Aug 1;189(2):313–335. doi: 10.1042/bj1890313

Physicochemical characterization of the ribosomal RNA species of the Mollusca. Molecular weight, integrity and secondary-structure features of the RNA of the large and small ribosomal subunits.

P Cammarano, P Londei, F Mazzei, A Felsani
PMCID: PMC1162002  PMID: 7458915

Abstract

1. The rRNA species of the Cephalopoda Octopus vulgaris and Loligo vulgaris were found to have unexpectedly high sedimentation coefficients and molecular weights. In 0.1 M-NaCl the L-rRNA (RNA from large ribosomal subunit) has the same s20 value as the L-rRNA of the mammals (30.7S), whereas the S-rRNA (RNA from small ribosomal subunit) sediments at a faster rate (20.1S) than the S-rRNA of both the mammals and the fungi (Neurospora crassa) (17.5S). The molecular weights of the L-rRNA were determined by gel electrophoresis in formamide and found to be 1.66 X 10(6) (Octupus) and 1.89 X 10(6) (Loligo); the mol.wt. of the S-rRNA of both species is 0.96 X 10(6), i.e. much larger than that of the mammals (0.65 X 10(6)) and almost coincident with that of the '23S' RNA of the prokaryotes. 2. By contrast, the less evolved Gastropoda and Lamellibranchiata (Murex trunculus and Macrocallista chione) have S-rRNA and L-rRNA species with mol.wts. of 0.65 X 10(6) and approx. 1.40 X 10(6).3. All the mature L-rRNA molecules of the cephalopoda are composed of two unequal fragments held together by regions of hydrogen-bonding having a similar, low, thermal stability in the two species; the molecular weights of the two fragments composing the L-rRNA are estimated to be 0.96 X 10(6) and 0.88 X 10(6) (Loligo) and 0.96 X 10(6) and 0.65 X 10(6) (Octupus). THe S-rRNA of both species is a continuous chain with exactly the same molecular weight (0.96 X 10(6)) as the heavier of the two fragments of the L-rRNA. 4. The secondary-structure features of the L-rRNA and S-rRNA species of the Caphalopoda were investigated by thermal 'melting' analysis in 4.0 M-guanidinium chloride; 60-70% of the residues are estimated to form short, independently 'melting' bihelical segments not more than 10 base-pairs in length. 5. Bases are unevenly distributed between non-helical and bihelical portions of the rRNA molecules, G and C residues being preferentially concentrated in bihelical comains. 6. The secondary-structure regions of the L-rRNA species of Octopus and Loligo are heterogenous, including two discrete fractions of independently 'melting' species that give rise to biphasic 'melting' profiles: a fraction consisting of shorter (G + C)-poorer segments (60-68% G + C, not more than 5 base-pairs in length) and a fraction consisting of longer (G + C)-richer segments (80-88% G + C, 5-10 base-pairs in length). No evidence for heterogeneity has been detected in the S-rRNa.

Full text

PDF
313

Selected References

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

  1. BRUNNGRABER E. F. A simplified procedure for the preparation of "soluble" RNA from rat liver. Biochem Biophys Res Commun. 1962 Jun 19;8:1–3. doi: 10.1016/0006-291x(62)90223-1. [DOI] [PubMed] [Google Scholar]
  2. Baralle F. E. Complete nucleotide sequence of the 5' noncoding region of rabbit beta-globin mRNA. Cell. 1977 Apr;10(4):549–558. doi: 10.1016/0092-8674(77)90088-5. [DOI] [PubMed] [Google Scholar]
  3. Boedtker H. The reaction of ribonucleic acid with formaldehyde. I. Optical absorbance studies. Biochemistry. 1967 Sep;6(9):2718–2727. doi: 10.1021/bi00861a011. [DOI] [PubMed] [Google Scholar]
  4. Cammarano P., Felsani A., Romeo A., Alberghina F. M. Particle weights of active ribosomal subunits from Neurospora crassa. Biochim Biophys Acta. 1973 May 18;308(3):404–411. doi: 10.1016/0005-2787(73)90333-x. [DOI] [PubMed] [Google Scholar]
  5. Cammarano P., Pons S., Londei P. Discontinuity of the large ribosomal subunit RNA and rRNA molecular weights in eukaryote evolution. Acta Biol Med Ger. 1975;34(7):1123–1135. [PubMed] [Google Scholar]
  6. Cox R. A. A spectrophotometric study of the secondary structure of ribonucleic acid isolated from the smaller and larger ribosomal subparticles of rabbit reticulocytes. Biochem J. 1970 Mar;117(1):101–118. doi: 10.1042/bj1170101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox R. A., Godwin E., Hastings J. R. Spectroscopic evidence for the uneven distribution of adenine and uracil residues in ribosomal ribonucleic acid of Drosophila melanogaster and of Plasmodium knowlesi and its possible evolutionary significance. Biochem J. 1976 Jun 1;155(3):465–475. doi: 10.1042/bj1550465a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cox R. A., Kanagalingam K. A spectrophotometric study of the secondary structure of ribonucleic acid based on a method for diminishing single-stranded base-'stacking' without affecting multi-helical structures. Biochem J. 1967 Jun;103(3):749–758. doi: 10.1042/bj1030749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dasgupta R., Shih D. S., Saris C., Kaesberg P. Nucleotide sequence of a viral RNA fragment that binds to eukaryotic ribosomes. Nature. 1975 Aug 21;256(5519):624–628. doi: 10.1038/256624a0. [DOI] [PubMed] [Google Scholar]
  10. Dawid I. B., Chase J. W. Mitochondrial RNA in Xenopus laevis. II. Molecular weights and other physical properties of mitochondrial ribosomal and 4 s RNA. J Mol Biol. 1972 Jan 28;63(2):217–231. doi: 10.1016/0022-2836(72)90371-3. [DOI] [PubMed] [Google Scholar]
  11. Ehresmann C., Stiegler P., Mackie G. A., Zimmermann R. A., Ebel J. P., Fellner P. Primary sequence of the 16S ribosomal RNA of Escherichia coli. Nucleic Acids Res. 1975 Feb;2(2):265–278. doi: 10.1093/nar/2.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gould H. J., Hamlyn P. H. The molecular weight of rabbit globin messenger RNA's. FEBS Lett. 1973 Mar 15;30(3):301–304. doi: 10.1016/0014-5793(73)80674-x. [DOI] [PubMed] [Google Scholar]
  13. Granboulan N., Scherrer K. Visualisation in the electron microscope and size of RNA from animal cells. Eur J Biochem. 1969 May 1;9(1):1–20. doi: 10.1111/j.1432-1033.1969.tb00569.x. [DOI] [PubMed] [Google Scholar]
  14. Kallenbach N. R. Theory of thermal transitions in low molecular weight RNA chains. J Mol Biol. 1968 Nov 14;37(3):445–466. doi: 10.1016/0022-2836(68)90114-9. [DOI] [PubMed] [Google Scholar]
  15. King H. W., Gould H. Low molecular weight ribonucleic acid in rabbit reticulocyte ribosomes. J Mol Biol. 1970 Aug;51(3):687–702. doi: 10.1016/0022-2836(70)90017-3. [DOI] [PubMed] [Google Scholar]
  16. Lava-Sanchez P. A., Amaldi F., Posta A. L. Base composition of ribosomal RNA and evolution. J Mol Evol. 1972 Dec 29;2(1):44–55. doi: 10.1007/BF01653942. [DOI] [PubMed] [Google Scholar]
  17. Lava-Sanchez P. A., Puppo S. Occurrence in vivo of "hidden breaks" at specific sites of 26 S ribosomal RNA of Musca carnaria. J Mol Biol. 1975 Jun 15;95(1):9–20. doi: 10.1016/0022-2836(75)90331-9. [DOI] [PubMed] [Google Scholar]
  18. Legon S. Characterization of the ribosome-protected regions of 125I-labelled rabbit globin messenger RNA. J Mol Biol. 1976 Sep 5;106(1):37–53. doi: 10.1016/0022-2836(76)90299-0. [DOI] [PubMed] [Google Scholar]
  19. Loening U. E. Molecular weights of ribosomal RNA in relation to evolution. J Mol Biol. 1968 Dec;38(3):355–365. doi: 10.1016/0022-2836(68)90391-4. [DOI] [PubMed] [Google Scholar]
  20. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  21. MATTHAEI J. H., NIRENBERG M. W. Characteristics and stabilization of DNAase-sensitive protein synthesis in E. coli extracts. Proc Natl Acad Sci U S A. 1961 Oct 15;47:1580–1588. doi: 10.1073/pnas.47.10.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pene J. J., Knight E., Jr, Darnell J. E., Jr Characterization of a new low molecular weight RNA in HeLa cell ribosomes. J Mol Biol. 1968 May 14;33(3):609–623. doi: 10.1016/0022-2836(68)90309-4. [DOI] [PubMed] [Google Scholar]
  23. Petermann M. L., Pavlovec A. The subunits and structural ribonucleic acids of Jensen sarcoma ribosomes. Biochim Biophys Acta. 1966 Feb 21;114(2):264–276. doi: 10.1016/0005-2787(66)90308-x. [DOI] [PubMed] [Google Scholar]
  24. Pinder J. C., Staynov D. Z., Gratzer W. B. Electrophoresis of RNA in formamide. Biochemistry. 1974 Dec 17;13(26):5373–5378. doi: 10.1021/bi00723a019. [DOI] [PubMed] [Google Scholar]
  25. Rawson J. R., Stutz E. Characterization of Euglena cytoplasmic ribosomes and ribosomal RNA by zone velocity sedimentation IN SUCROSE GRADIENTS. J Mol Biol. 1968 Apr 14;33(1):309–314. doi: 10.1016/0022-2836(68)90296-9. [DOI] [PubMed] [Google Scholar]
  26. Rungger D., Crippa M. The primary ribosomal DNA transcript in eukaryotes. Prog Biophys Mol Biol. 1977;31(3):247–269. doi: 10.1016/0079-6107(78)90010-x. [DOI] [PubMed] [Google Scholar]
  27. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  28. Shine J., Dalgarno L. Occurrence of heat-dissociable ribosomal RNA in insects: the presence of three polynucleotide chains in 26 S RNA from cultured Aedes aegypti cells. J Mol Biol. 1973 Mar 25;75(1):57–72. doi: 10.1016/0022-2836(73)90528-7. [DOI] [PubMed] [Google Scholar]
  29. Stevens A. R., Pachler P. F. Discontinuity of 26 s rRNA in Acanthamoeba castellani. J Mol Biol. 1972 May 14;66(2):225–237. doi: 10.1016/0022-2836(72)90475-5. [DOI] [PubMed] [Google Scholar]
  30. Stutz E., Noll H. Characterization of cytoplasmic and chloroplast polysomes in plants: evidence for three classes of ribosomal RNA in nature. Proc Natl Acad Sci U S A. 1967 Mar;57(3):774–781. doi: 10.1073/pnas.57.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sy J., McCarty K. S. Characterization of 5.8-S RNA from a complex with 26-S ribosomal RNA from Arbacia punctulata. Biochim Biophys Acta. 1970 Jan 21;199(1):86–94. doi: 10.1016/0005-2787(70)90697-0. [DOI] [PubMed] [Google Scholar]
  32. Van de Voorde A., Contreras R., Rogiers R., Fiers W. The initiation region of the SV40 VP1 gene. Cell. 1976 Sep;9(1):117–120. doi: 10.1016/0092-8674(76)90057-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES