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. 1959 Jan 25;5(1):1–10. doi: 10.1083/jcb.5.1.1

A Cytochemical Study on the Pancreas of the Guinea Pig

IV. Chemical and Metabolic Investigation of the Ribonucleoprotein Particles

Philip Siekevitz 1, George E Palade 1
PMCID: PMC2224619  PMID: 13630927

Abstract

Five ribonucleoprotein (RNP) fractions were isolated from the postmitochondrial supernatant of the pancreas of the guinea pig. Two were obtained from the microsomes which, by deoxycholate (DOC) treatment, were subdivided into a DOC-soluble and a DOC-insoluble fraction. The latter was taken to represent attached RNP particles. Two other fractions obtained from the microsomal supernatant supposedly represent free RNP particles existing as such in the cytoplasm, while a third fraction resisted sedimentation for 20 hours at 105,000 g and is considered to be a soluble nucleoprotein. These fractions exhibited different RNA/protein ratios and also different RNA turnover patterns, as determined after in vivo labelling with adenine-8-C14. However, little discernible differences could be detected in the nucleotide composition of the RNA moieties of these RNP fractions. Amino acid-"activating" enzymes were found to occur in the fraction containing the soluble nucleoproteins. The discussion focuses on the relationships between these fractions and protein synthesis in the pancreas, using data given in this and a previous paper, and data contained in the literature.

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Selected References

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

  1. BARNUM C. P., HUSEBY R. A., VERMUND H. A time study of the incorporation of radiophosphorus into the nucleic acids and other compounds of a transplanted mouse mammary carcinoma. Cancer Res. 1953 Dec;13(12):880–889. [PubMed] [Google Scholar]
  2. BENNETT E. L. Incorporation of adenine into nucleotides and nucleic acids of C57 mice. Biochim Biophys Acta. 1953 Aug;11(4):487–496. doi: 10.1016/0006-3002(53)90086-9. [DOI] [PubMed] [Google Scholar]
  3. BHARGAVA P. M., SIMKIN J. L., WORK T. S. Incorporation of radioactive phosphorus into the ribonucleic acid of subfractions derived from guinea-pig-liver microsomes. Biochem J. 1958 Feb;68(2):265–269. doi: 10.1042/bj0680265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CANELLAKIS E. S. On the mechanism of incorporation of adenylic acid form adenosine triphosphate into ribonucleic acid by soluble mammalian enzyme systems. Biochim Biophys Acta. 1957 Jul;25(1):217–218. [PubMed] [Google Scholar]
  5. CHAUVEAU J., MOULE Y., ROUILLER C. Localisation de l'acide ribonucléique dans les diverses structures morphologiques des microsomes de foie de rat. Exp Cell Res. 1957 Oct;13(2):398–400. doi: 10.1016/0014-4827(57)90019-8. [DOI] [PubMed] [Google Scholar]
  6. COLE R. D., COOTE J., WORK T. S. Activation of amino-acids by soluble enzymes from pancreas and other tissues. Nature. 1957 Jan 26;179(4552):199–199. doi: 10.1038/179199a0. [DOI] [PubMed] [Google Scholar]
  7. DAVIE E. W., KONINGSBERGER V. V., LIPMANN F. The isolation of a tryptophan-activating enzyme from pancreas. Arch Biochem Biophys. 1956 Nov;65(1):21–38. doi: 10.1016/0003-9861(56)90173-4. [DOI] [PubMed] [Google Scholar]
  8. DE LAMIRANDE G., ALLARD C., CANTERO A. Heterogeneity of intracellular ribonucleic acid in rat liver cells. J Biol Chem. 1955 Jun;214(2):519–524. [PubMed] [Google Scholar]
  9. DEMOSS J. A., NOVELLI G. D. An amino acid dependent exchange between 32P labeled inorganic pyrophosphate and ATP in microbial extracts. Biochim Biophys Acta. 1956 Oct;22(1):49–61. doi: 10.1016/0006-3002(56)90222-0. [DOI] [PubMed] [Google Scholar]
  10. EDMONDS M., ABRAMS R. Incorporation of ATP into polynucleotide in extracts of Ehrlich ascites cells. Biochim Biophys Acta. 1957 Oct;26(1):226–227. doi: 10.1016/0006-3002(57)90086-0. [DOI] [PubMed] [Google Scholar]
  11. HECHT L. I., POTTER V. R., TAKAGI Y. Nucleic acid metabolism in regenerating rat liver. II. Studies on growing rats. Cancer Res. 1956 Nov;16(10 Pt 1):994–998. [PubMed] [Google Scholar]
  12. HERBERT E., POTTER V. R., HECHT L. I. Nucleotide metabolism. VII. The incorporation of radioactivity from orotic acid-6-C14 into ribonucleic acid in cell-free systems from rat liver. J Biol Chem. 1957 Apr;225(2):659–674. [PubMed] [Google Scholar]
  13. HERBERT E. The incorporation of adenine nucleotides into ribonucleic acid of cell-free systems from liver. J Biol Chem. 1958 Apr;231(2):975–986. [PubMed] [Google Scholar]
  14. HOAGLAND M. B., KELLER E. B., ZAMECNIK P. C. Enzymatic carboxyl activation of amino acids. J Biol Chem. 1956 Jan;218(1):345–358. [PubMed] [Google Scholar]
  15. HOAGLAND M. B., STEPHENSON M. L., SCOTT J. F., HECHT L. I., ZAMECNIK P. C. A soluble ribonucleic acid intermediate in protein synthesis. J Biol Chem. 1958 Mar;231(1):241–257. [PubMed] [Google Scholar]
  16. HUSEBY R. A., BARNUM C. P. Investigation of the phosphorus-containing constituents of centrifugally prepared fractions from mouse liver cell cytoplasm. Arch Biochem. 1950 Apr;26(2):187–198. [PubMed] [Google Scholar]
  17. JARDETZKY C. D., BARNUM C. P. Metabolic activity and intracellular distribution of nucleic acid phosphorus in regenerating liver. Arch Biochem Biophys. 1957 Apr;67(2):350–365. doi: 10.1016/0003-9861(57)90290-4. [DOI] [PubMed] [Google Scholar]
  18. Jorpes E. On the pentose polynucleotide of the pancreas. Biochem J. 1934;28(6):2102–2108. doi: 10.1042/bj0282102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. KEMP J. W., ALLEN F. W. Ribonucleic acids from pancreas which contain new components. Biochim Biophys Acta. 1958 Apr;28(1):51–58. doi: 10.1016/0006-3002(58)90426-8. [DOI] [PubMed] [Google Scholar]
  20. KERR S. E., SERAIDARIAN K., WARGON M. On the composition of the ribonucleic acid of beef pancreas, with notes on the action of ribonuclease. J Biol Chem. 1949 Dec;181(2):773–780. [PubMed] [Google Scholar]
  21. KIRBY K. S. A new method for the isolation of ribonucleic acids from mammalian tissues. Biochem J. 1956 Nov;64(3):405–408. doi: 10.1042/bj0640405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. LAIRD A. K., BARTON A. D., NYGAARD O. Synthesis of protein and ribonucleic acid in rat liver during refeeding after starvation. Exp Cell Res. 1955 Dec;9(3):523–540. doi: 10.1016/0014-4827(55)90081-1. [DOI] [PubMed] [Google Scholar]
  23. Lipmann F. Chairman's Introduction: Some Facts and Problems. Proc Natl Acad Sci U S A. 1958 Feb;44(2):67–73. doi: 10.1073/pnas.44.2.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MARSHAK A. Purine and pyrimidine content of the nucleic acids of nuclei and cytoplasm. J Biol Chem. 1951 Apr;189(2):607–615. [PubMed] [Google Scholar]
  25. NISMANN B., BERGMANN F. H., BERG P. Observations on amino acid-dependent exchanges of inorganic pyrophosphate and ATP. Biochim Biophys Acta. 1957 Dec;26(3):639–640. doi: 10.1016/0006-3002(57)90113-0. [DOI] [PubMed] [Google Scholar]
  26. NYGAARD O., RUSCH H. P. Incorporation of radioactive phosphate into nucleic acids of regenerating rat liver. Cancer Res. 1955 May;15(4):240–245. [PubMed] [Google Scholar]
  27. OGATA K., NOHARA H. The possible role of the ribonucleic acid (RNA) of the pH 5 enzyme in amino acid activation. Biochim Biophys Acta. 1957 Sep;25(3):659–660. doi: 10.1016/0006-3002(57)90549-8. [DOI] [PubMed] [Google Scholar]
  28. OLMSTED P. S., VILLEE C. A. Nucleic acid composition of human liver cell fractions. J Biol Chem. 1955 Jan;212(1):179–186. [PubMed] [Google Scholar]
  29. PALADE G. E. Intracisternal granules in the exocrine cells of the pancreas. J Biophys Biochem Cytol. 1956 Jul 25;2(4):417–422. doi: 10.1083/jcb.2.4.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. PALADE G. E., SIEKEVITZ P. Liver microsomes; an integrated morphological and biochemical study. J Biophys Biochem Cytol. 1956 Mar 25;2(2):171–200. doi: 10.1083/jcb.2.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. PARDEE A. B., PRESTIDGE L. S. The dependence of nucleic acid synthesis on the presence of amino acids in Escherichia coli. J Bacteriol. 1956 Jun;71(6):677–683. doi: 10.1128/jb.71.6.677-683.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. REID E., STEVENS B. M. Functions of ribonucleic acid in liver cytoplasm. Biochim Biophys Acta. 1956 Mar;19(3):554–555. doi: 10.1016/0006-3002(56)90484-x. [DOI] [PubMed] [Google Scholar]
  33. SACKS J., HURLEY P. D., YOUNG J. M., Jr The metabolic inhomogeneity of ribonucleic acid phosphorus. J Biol Chem. 1955 Jun;214(2):723–730. [PubMed] [Google Scholar]
  34. SACKS J., SAMARTH K. D. Metabolic inhomogeneity in ribonucleic acid phosphorus of particulate fractions of liver. J Biol Chem. 1956 Nov;223(1):423–429. [PubMed] [Google Scholar]
  35. SCHWEET R. S., HOLLEY R. W., ALLEN E. H. Amino acid activation in hog pancreas. Arch Biochem Biophys. 1957 Oct;71(2):311–325. doi: 10.1016/0003-9861(57)90040-1. [DOI] [PubMed] [Google Scholar]
  36. SHIGEURA H. T., CHARGAFF E. Comparative studies of protein and ribonucleic acid formation. Biochim Biophys Acta. 1957 May;24(2):450–451. doi: 10.1016/0006-3002(57)90231-7. [DOI] [PubMed] [Google Scholar]
  37. SIEKEVITZ P., PALADE G. E. A cyto-chemical study on the pancreas of the guinea pig. III. In vivo incorporation of leucine-1-C14 into the proteins of cell fractions. J Biophys Biochem Cytol. 1958 Sep 25;4(5):557–566. doi: 10.1083/jcb.4.5.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. SIEKEVITZ P., PALADE G. E. A cytochemical study on the pancreas of the guinea pig. I. Isolation and enzymatic activities of cell fractions. J Biophys Biochem Cytol. 1958 Mar 25;4(2):203–218. doi: 10.1083/jcb.4.2.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. SIEKEVITZ P., PALADE G. E. A cytochemical study on the pancreas of the guinea pig. II. Functional variations in the enzymatic activity of microsomes. J Biophys Biochem Cytol. 1958 May 25;4(3):309–318. doi: 10.1083/jcb.4.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. SIEKEVITZ P. Uptake of radioactive alanine in vitro into the proteins of rat liver fractions. J Biol Chem. 1952 Apr;195(2):549–565. [PubMed] [Google Scholar]
  41. SIMKIN J. L., WORK T. S. Incorporation of radioactive amino acids into proteins of the microsome fraction of guinea-pig liver in a cell-free system. Biochem J. 1957 Dec;67(4):617–624. doi: 10.1042/bj0670617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. SMELLIE R. M., MCINDOE W. M., LOGAN R., DAVIDSON J. N., DAWSON I. M. Phosphorus compounds in the cell. IV. The incorporation of radioactive phosphorus into liver cell fractions. Biochem J. 1953 May;54(2):280–286. doi: 10.1042/bj0540280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. SOYENKOFF B. C. An improved micromethod of phosphate determination. J Biol Chem. 1952 Sep;198(1):221–227. [PubMed] [Google Scholar]
  44. Schweet R. S., Bovard F. C., Allen E., Glassman E. THE INCORPORATION OF AMINO ACIDS INTO RIBONUCLEIC ACID. Proc Natl Acad Sci U S A. 1958 Feb;44(2):173–177. doi: 10.1073/pnas.44.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. TYNER E. P., HEIDELBERGER C., LEPAGE G. A. Intracellular distribution of radioactivity in nucleic acid nucleotides and proteins following simultaneous administration of P32 and glycine-2-C14. Cancer Res. 1953 Feb;13(2):186–203. [PubMed] [Google Scholar]
  46. WEBSTER G. C. Amino acid incorporation by intact and disrupted ribonucleoprotein particles. J Biol Chem. 1957 Nov;229(1):535–546. [PubMed] [Google Scholar]
  47. Weiss S. B., Acs G., Lipmann F. AMINO ACID INCORPORATION IN PIGEON PANCREAS FRACTIONS. Proc Natl Acad Sci U S A. 1958 Feb;44(2):189–196. doi: 10.1073/pnas.44.2.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zamecnik P. C., Stephenson M. L., Hecht L. I. Intermediate Reactions in Amino Acid Incorporation. Proc Natl Acad Sci U S A. 1958 Feb;44(2):73–78. doi: 10.1073/pnas.44.2.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zioudrou C., Fujii S., Fruton J. S. LABELING OF PROTEINS BY ISOTOPIC AMINO ACID DERIVATIVES. Proc Natl Acad Sci U S A. 1958 May;44(5):439–446. doi: 10.1073/pnas.44.5.439. [DOI] [PMC free article] [PubMed] [Google Scholar]

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