Abstract
Two populations of polyribosomes have been isolated from third instar larvae of D. melanogaster. One population appeared to be soluble while the second seemed membrane-bound. Short-term labeling of the two RNP fractions with radioactive nucleic acid and protein precursors was achieved by using a feeding stimulant. RNA was extracted from both polyribosomal fractions following 25, 40, and 60 min of in vivo uridine-3H incorporation. Soluble polyribosomes exhibited more rapid uptake of uridine into ribosomal and heterogeneous RNA fractions than did membrane-bound polyribosomes at comparable time periods. In vivo amino acid incorporation into the two polyribosomal populations was examined after 10, 20, 40, 60, and 80 min of incubation in leucine-3H. In this case, the membrane-bound polyribosomes reached a higher specific activity than did the soluble ones. These functional differences confirmed the observation, based on cellular fractionation studies, that the two classes of polyribosomes represented functionally distinct populations. These data have been compared with those from studies on other metazoan systems. In addition, dithiothreitol has been demonstrated to be a powerful ribonuclease inhibitor.
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- BROWN D. D., LITTNA E. VARIATIONS IN THE SYNTHESIS OF STABLE RNA'S DURING OOGENESIS AND DEVELOPMENT OF XENOPUS LAEVIS. J Mol Biol. 1964 May;8:688–695. doi: 10.1016/s0022-2836(64)80117-0. [DOI] [PubMed] [Google Scholar]
- Brown D. D., Gurdon J. B. Size distribution and stability of DNA-like RNA synthesized during development of anucleolate embryos of Xenopus laevis. J Mol Biol. 1966 Aug;19(2):399–422. doi: 10.1016/s0022-2836(66)80013-x. [DOI] [PubMed] [Google Scholar]
- Darnell J. E., Jr Ribonucleic acids from animal cells. Bacteriol Rev. 1968 Sep;32(3):262–290. doi: 10.1128/br.32.3.262-290.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GIRARD M., LATHAM H., PENMAN S., DARNELL J. E. ENTRANCE OF NEWLY FORMED MESSENGER RNA AND RIBOSOMES INTO HELA CELL CYTOPLASM. J Mol Biol. 1965 Feb;11:187–201. doi: 10.1016/s0022-2836(65)80050-x. [DOI] [PubMed] [Google Scholar]
- Goldberg B., Green H. Collagen synthesis on polyribosomes of cultured mammalian fibroblasts. J Mol Biol. 1967 May 28;26(1):1–18. doi: 10.1016/0022-2836(67)90257-4. [DOI] [PubMed] [Google Scholar]
- Greenberg J. R. Synthesis and properties of ribosomal RNA in Drosophila. J Mol Biol. 1969 Nov 28;46(1):85–98. doi: 10.1016/0022-2836(69)90059-x. [DOI] [PubMed] [Google Scholar]
- HORIKAWA M., FOX A. S. CULTURE OF EMBRYONIC CELLS OF DROSOPHILA MELANOGASTER IN VITRO. Science. 1964 Sep 25;145(3639):1437–1439. doi: 10.1126/science.145.3639.1437. [DOI] [PubMed] [Google Scholar]
- Hallinan T., Munro H. N. Protein synthesis and ribonucleic acid turnover in rat-liver microsome subfractions. Biochim Biophys Acta. 1965 Oct 11;108(2):285–296. doi: 10.1016/0005-2787(65)90013-4. [DOI] [PubMed] [Google Scholar]
- Hastings J. R., Kirby K. S. The nucleic acids of Drosophila melanogaster. Biochem J. 1966 Aug;100(2):532–539. doi: 10.1042/bj1000532. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hennig W. Ribonucleic acid synthesis of the Y-chromosome of Drosophila hydei. J Mol Biol. 1968 Dec 14;38(2):227–239. doi: 10.1016/0022-2836(68)90408-7. [DOI] [PubMed] [Google Scholar]
- Henshaw E. C., Revel M., Hiatt H. H. A cytoplasmic particle bearing messenger ribonucleic acid in rat liver. J Mol Biol. 1965 Nov;14(1):241–256. doi: 10.1016/s0022-2836(65)80244-3. [DOI] [PubMed] [Google Scholar]
- Joklik W. K., Becker Y. Studies on the genesis of polyribosomes. I. Origin and significance of the subribosomal particles. J Mol Biol. 1965 Sep;13(2):496–510. doi: 10.1016/s0022-2836(65)80112-7. [DOI] [PubMed] [Google Scholar]
- Joklik W. K., Becker Y. Studies on the genesis of polyribosomes. II. The association of nascent messenger RNA with the 40 S subribosomal particle. J Mol Biol. 1965 Sep;13(2):511–520. doi: 10.1016/s0022-2836(65)80113-9. [DOI] [PubMed] [Google Scholar]
- Latham H., Darnell J. E. Distribution of mRNA in the cytoplasmic polyribosomes of the HeLa cell. J Mol Biol. 1965 Nov;14(1):1–12. doi: 10.1016/s0022-2836(65)80224-8. [DOI] [PubMed] [Google Scholar]
- 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]
- Martin T. E., Rolleston F. S., Low R. B., Wool I. G. Dissociation and reassociation of skeletal muscle ribosomes. J Mol Biol. 1969 Jul 14;43(1):135–149. doi: 10.1016/0022-2836(69)90084-9. [DOI] [PubMed] [Google Scholar]
- Noll H. Characterization of macromolecules by constant velocity sedimentation. Nature. 1967 Jul 22;215(5099):360–363. doi: 10.1038/215360a0. [DOI] [PubMed] [Google Scholar]
- 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]
- RABINOWITZ M., ZAK R., BELLER B., RAMPERSAD O., WOOL I. G. EFFECT OF PROTEOLYTIC ENZYMES ON SEDIMENTATION PROPERTIES OF RIBONUCLEOPROTEIN PARTICLES FROM HEART MUSCLE. Proc Natl Acad Sci U S A. 1964 Dec;52:1353–1360. doi: 10.1073/pnas.52.6.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RIFKIND R. A., LUZZATTO L., MARKS P. A. SIZE OF POLYRIBOSOMES IN INTACT RETICULOCYTES. Proc Natl Acad Sci U S A. 1964 Nov;52:1227–1232. doi: 10.1073/pnas.52.5.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RITOSSA F. M., SPIEGELMAN S. LOCALIZATION OF DNA COMPLEMENTARY TO RIBOSOMAL RNA IN THE NUCLEOLUS ORGANIZER REGION OF DROSOPHILA MELANOGASTER. Proc Natl Acad Sci U S A. 1965 Apr;53:737–745. doi: 10.1073/pnas.53.4.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Redman C. M., Sabatini D. D. Vectorial discharge of peptides released by puromycin from attached ribosomes. Proc Natl Acad Sci U S A. 1966 Aug;56(2):608–615. doi: 10.1073/pnas.56.2.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ritossa F. M., Atwood K. C., Spiegelman S. A molecular explanation of the bobbed mutants of Drosophila as partial deficiencies of "ribosomal" DNA. Genetics. 1966 Sep;54(3):819–834. doi: 10.1093/genetics/54.3.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ritossa F. M., Atwood K. C. Unequal proportions of DNA complementary to ribosomal RNA in males and females of Drosophila simulans. Proc Natl Acad Sci U S A. 1966 Aug;56(2):496–499. doi: 10.1073/pnas.56.2.496. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sabatini D. D., Tashiro Y., Palade G. E. On the attachment of ribosomes to microsomal membranes. J Mol Biol. 1966 Aug;19(2):503–524. doi: 10.1016/s0022-2836(66)80019-0. [DOI] [PubMed] [Google Scholar]
- Talal N., Kaltreider H. B. Functional and structural studies of membrane-bound and free ribosomes from rat spleen. J Biol Chem. 1968 Dec 25;243(24):6504–6510. [PubMed] [Google Scholar]
- Vesco C., Penman S. The cytoplasmic RNA of HeLa cells: new discrete species associated with mitochondria. Proc Natl Acad Sci U S A. 1969 Jan;62(1):218–225. doi: 10.1073/pnas.62.1.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WARNER J. R., KNOPF P. M., RICH A. A multiple ribosomal structure in protein synthesis. Proc Natl Acad Sci U S A. 1963 Jan 15;49:122–129. doi: 10.1073/pnas.49.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WETTSTEIN F. O., STAEHELIN T., NOLL H. Ribosomal aggregate engaged in protein synthesis: characterization of the ergosome. Nature. 1963 Feb 2;197:430–435. doi: 10.1038/197430a0. [DOI] [PubMed] [Google Scholar]
