Abstract
Nucleic acid reassociation methods were used to estimate the number of different polyadenylated RNA [poly (A) + RNA] sequences in the cytoplasm of whole Drosophila melanogaster at different stages of development and in the cytoplasm of cells of the L3 cell line. The number-average length (LN) of poly (A) + RNA from L3 cells is 1.4 kb, and the steady-state LN of the poly (A) tracts is 70 nucleotides. Analysis of RNA-driven reassociation with copy DNA shows that the poly (A) + RNA from L3 cells contains 5200 different sequences distributed in three abundance classes. The RNA forms hybrid duplexes with about 4.5% of single-copy Drosophila DNA, corresponding to 6500 sequences of 1.4 kb. The LN of poly (A) tracts present in whole embryos, larvae, pupae, and imagos is in each case close to 70 nucleotides. RNA-driven reassociation experiments show that poly (A) + RNA from these sources contains, respectively, 3500,≥4900, 6900, and ≥4900 sequences. Cross-hybridization reactions show extensive homology between these RNA populations. All five poly (A) + RNA preparations contain a prominent component with a sedimentation coefficient of 13 and a size of 1.78 kb, which is shown to be the larger species of mitochondrial rRNA. This rRNA binds to oligo (dT) cellulose and serves as an efficient template for cDNA synthesis by reverse transcriptase. In RNA-driven reassociation experiments it behaves like an abundant mRNA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Axel, R., Feigelson, P., and Schutz, G. (1976). Analysis of the complexity and diversity of mRNA from chicken liver and oviduct. Cell 7247.
Birnie, G. D., Macphail, E., Young, B. D., Getz, M. J., and Paul, J. (1974). The diversity of the messenger RNA population in growing Friend cells. Cell Differ. 3221.
Bishop, J. O. (1974). The gene numbers game. Cell 281.
Bishop, J. O., and Rosbash, M. (1973). Reiteration frequency of duck haemoglobin genes. Nature New Biol. 241204.
Bishop, J. O., Morton, J. G., Rosbash, M., and Richardson, M. (1974a). Three abundance classes in HeLa cell mRNA. Nature 250199.
Bishop, J. O., Rosbash, M., and Evans, D. (1974b). Polynucleotide sequences in eukaryotic DNA and RNA that form ribonuclease-resistant complexes with poly(U). J. Mol. Biol. 85:75.
Bishop, J. O., Beckmann, J. S., Campo, M. S., Hastie, N. D., Izquierdo, M., and Perlman, S. (1975). DNA-RNA hybridisation. Phil. Tr. Roy. Soc. Lond. Ser. B 272147.
Bultmann, H., and Laird, C. D. (1973). Mitochondrial DNA from Drosophila melanogaster. Biochim. Biophys. Acta 299196.
Campo, M. S., and Bishop, J. O. (1974). Two classes of mRNA in cultured rat cells: repetitive sequence transcripts and unique sequence transcripts. J. Mol. Biol. 90:649.
Davidson, E. H., and Hough, B. R. (1971). Genetic information in oocyte RNA. J. Mol. Biol. 56491.
Gierer, A. (1958). Vergleichende Untersuchungen an hochmolekularer Ribosenucleinsaure. Z. Naturforsch. 13 788.
Goldberg, R. B., Galau, G., Britten, R. J., and Davidson, E. H. (1973). Nonrepetitive DNA sequence representation in sea urchin mRNA. Proc. Natl. Acad. Sci. 703516.
Hastie, N. D., and Bishop, J. O. (1976). The expression of three abundance classes of mRNA in mouse tissues. Cell 9761.
Kaufmann, Y., Milcarek, C., Berissi, H., and Penman, S. (1977). HeLa cell poly(A)−mRNA codes for a subset of poly(A)+RNA-directed proteins with an actin as a major product. Proc. Natl. Acad. Sci. 744801.
Klukas, C. K. and Dawid, I. B. (1976). Characterisation and mapping of mitochondrial ribosomal RNA and mitochondrial DNA in Drosophila melanogaster. Cell 9615.
Lefevre, G. (1974). The relationship between genes and polytene chromosome bands. Ann. Rev. Genet. 851.
Lefevre, G. (1976). A photographic representation and interpretation of the polytene chromosomes of Drosophila melanogaster salivary glands. In Ashburner, M., and Novitsky, E. (eds.), The Genetics and Biology of Drosophila, Vol. 1a, Academic Press, New York, pp. 31–66.
Levy, W. B., and McCarthy, B. J. (1975). Messenger RNA complexity in Drosophila melanogaster. Biochemistry 142440.
McKenzie, S., Henikoff, S., and Meselson, M. (1975). Localization of RNA from heat-induced polysomes at puff sites in Drosophila melanogaster. Proc. Natl. Acad. Sci. 721117.
Milcarek, C., Price, R., and Penman, S. (1974). The metabolism of a poly(A) minus mRNA fraction in HeLa cells. Cell 31.
Nemer, M., Graham, M., and Dubroff, L. M. (1974). Co-existence of nonhistone mRNA species lacking and containing polyadenylic acid in sea urchin embryos. J. Mol. Biol. 89435.
O'Farrell, P. H. (1975). High resolution two-dimensional separation of proteins. J. Biol. Chem. 2504007.
Paterson, B. M., and Bishop, J. O. (1977). Changes in the mRNA population of chick myoblasts during myogenesis in vitro. Cell 12751.
Peacock, W. J., Brutlag, D., Goldring, E., Appels, R., Hinton, C. W., and Lindsley, D. L. (1973). The organisation of highly repeated sequences in Drosophila melanogaster chromosomes. Cold Spring Harbor Symp. Quant. Biol. 38405.
Perlman, S., Phillips, C., and Bishop, J. O. (1976). A study of foldback DNA. Cell 833.
Prensky, W., Steffenson, D. M., and Hughes, W. L. (1973). The use of iodinated RNA for gene localization. Proc. Natl. Acad. Sci. 701860.
Ritossa, F. (1962). A new puffing pattern induced by temperature shock and DNP in Drosophila. Experientia 18571.
Rosbash, M., and Ford, P. J. (1974). Polyadenylic-acid-containing RNA in Xenopus laevis oocytes. J. Mol. Biol. 8587.
Ross, J., Aviv, H., Scolnick, E., and Leder, P. (1972). In vitro synthesis of DNA complementary to purified rabbit globin mRNA. Proc. Natl. Acad. Sci. 69264.
Schneider, I. (1972). Cell lines derived from late embryonic stages of Drosophila melanogaster. J. Embryol. Exp. Morphol. 27353.
Sheiness, D. K., and Darnell, J. E. (1973). Polyadenylic acid segment in mRNA becomes shorter with age. Nature New Biol. 113265.
Shields, G., Dubendorfer, A., and Sang, J. H. (1975). Differentiation in vitro of larval cell types from early embryonic cells of Drosophila melanogaster. J. Embryol. Exp. Morphol. 33159.
Singer, R. H., and Penman, S. (1973). Messenger RNA in HeLa cells: Kinetics of formation and decay. J. Mol. Biol. 78321.
Southern, E. M. (1975). Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98503.
Spirin, A. S. (1963). Some problems concerning the macromolecular structure of ribonucleic acids. Prog. Nucl. Acid. Res. 1301.
Spradling, A., Penman, S., Campo, M. S., and Bishop, J. O. (1974). Repetitious and unique sequences in the heterogeneous nuclear and cytoplasmic messenger RNA of mammalian and insect cells. Cell 323.
Staynov, D. Z., Pinder, J. C., and Gratzer, W. B. (1972). Molecular weight determination of nucleic acids by gel electrophoresis in non-aqueous solution. Nature New Biol. 235108.
Studier, F. W. (1965). Sedimentation studies of the size and shape of DNA. J. Mol. Biol. 11373.
Tissieres, A., Mitchell, H. K., and Tracy, U. M. (1974). Protein synthesis in salivary glands of Drosophila melanogaster: Relation to chromosome puffs. J. Mol. Biol. 84389.
Author information
Authors and Affiliations
Additional information
This work was supported by the Science Research Council and the Cancer Research Campaign.
Rights and permissions
About this article
Cite this article
Izquierdo, M., Bishop, J.O. An analysis of cytoplasmic RNA populations in Drosophila melanogaster, Oregon R. Biochem Genet 17, 473–497 (1979). https://doi.org/10.1007/BF00498885
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00498885