Abstract
We have determined the fraction of polyadenylated cytoplasmic RNA from trout testis complementary to unique and repetitive DNA. Some 21% of the cDNA probe representative of this RNA population renatures with rapid kinetics, characteristics of repetitive sequences. The major proportion of the cDNA renatures with unique sequence DNA. Experiments with fractionated cDNA probes allow us to conclude that, in trout testis, the most abundant polyadenylated mRNAs are not preferentially transcribed from repetitive DNA, as it has shown to be the case in two eukaryotic cell lines. Treatment of trout testis nuclei with DNase I, under conditions in which 10% of the total DNA is digested, preferentially depletes the DNA of sequences being transcribed into polyadenylated mRNA. These data confirm the results of H. Weintraub and M. Groundine [(1976) Science 193, 848-856] and those of A. Garel and R. Axel [(1976) Proc. Natl. Acad. Sci. U.S.A. 73, 3966-3970] and suggest that the conformation of DNA in the active genes of chromatin is such that it is more susceptible to digestion by DNaseI.
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- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Axel R., Cedar H., Felsenfeld G. Synthesis of globin ribonucleic acid from duck-reticulocyte chromatin in vitro. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2029–2032. doi: 10.1073/pnas.70.7.2029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Axel R., Feigelson P., Schutz G. Analysis of the complexity and diversity of mRNA from chicken liver and oviduct. Cell. 1976 Feb;7(2):247–254. doi: 10.1016/0092-8674(76)90024-6. [DOI] [PubMed] [Google Scholar]
- Beatriz L. W., McCarthy B. J. Messenger RNA complexity in Drosophila melanogaster. Biochemistry. 1975 Jun 3;14(11):2440–2446. doi: 10.1021/bi00682a026. [DOI] [PubMed] [Google Scholar]
- Biessmann H., Gjerset R. A., Levy B., McCarthy B. J. Fidelity of chromatin transcription in vitro. Biochemistry. 1976 Oct 5;15(20):4356–4363. doi: 10.1021/bi00665a002. [DOI] [PubMed] [Google Scholar]
- Bishop J. O., Morton J. G., Rosbash M., Richardson M. Three abundance classes in HeLa cell messenger RNA. Nature. 1974 Jul 19;250(463):199–204. doi: 10.1038/250199a0. [DOI] [PubMed] [Google Scholar]
- Bishop J. O., Rosbash M. Reiteration frequency of duck haemoglobin genes. Nat New Biol. 1973 Feb 14;241(111):204–207. doi: 10.1038/newbio241204a0. [DOI] [PubMed] [Google Scholar]
- Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
- Campo M. S., Bishop J. O. Two classes of messenger RNA in cultured rat cells: repetitive sequence transcripts and unique sequence transcripts. J Mol Biol. 1974 Dec 25;90(4):649–663. doi: 10.1016/0022-2836(74)90530-0. [DOI] [PubMed] [Google Scholar]
- Crouse G. F., Fodor J. B., Doty P. In vitro transcription of chromatin in the presence of a mercurated nucleotide. Proc Natl Acad Sci U S A. 1976 May;73(5):1564–1567. doi: 10.1073/pnas.73.5.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davidson E. H., Britten R. J. Organization, transcription, and regulation in the animal genome. Q Rev Biol. 1973 Dec;48(4):565–613. doi: 10.1086/407817. [DOI] [PubMed] [Google Scholar]
- Davidson E. H., Hough B. R., Amenson C. S., Britten R. J. General interspersion of repetitive with non-repetitive sequence elements in the DNA of Xenopus. J Mol Biol. 1973 Jun 15;77(1):1–23. doi: 10.1016/0022-2836(73)90359-8. [DOI] [PubMed] [Google Scholar]
- Firtel R. A., Jacobson A., Lodish H. F. Isolation and hybridization kinetics of messenger RNA from Dictyostelium discoideum. Nat New Biol. 1972 Oct 25;239(95):225–228. doi: 10.1038/newbio239225a0. [DOI] [PubMed] [Google Scholar]
- Garel A., Axel R. Selective digestion of transcriptionally active ovalbumin genes from oviduct nuclei. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3966–3970. doi: 10.1073/pnas.73.11.3966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gedamu L., Dixon G. H. Purification and properties of biologically active rainbow trout testis protamine mRNA. J Biol Chem. 1976 Mar 10;251(5):1455–1463. [PubMed] [Google Scholar]
- Gilmour R. S., Paul J. Tissue-specific transcription of the globin gene in isolated chromatin. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3440–3442. doi: 10.1073/pnas.70.12.3440. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldberg R. B., Galau G. A., Britten R. J., Davidson E. H. Nonrepetitive DNA sequence representation in sea urchin embryo messenger RNA. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3516–3520. doi: 10.1073/pnas.70.12.3516. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottesfeld J. M., Murphy R. F., Bonner J. Structure of transcriptionally active chromatin. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4404–4408. doi: 10.1073/pnas.72.11.4404. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greenberg J. R., Perry R. P. Hybridization properties of DNA sequences directing the synthesis of messenger RNA and heterogeneous nuclear RNA. J Cell Biol. 1971 Sep;50(3):774–786. doi: 10.1083/jcb.50.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grouse L., Chilton M. D., McCarthy B. J. Hybridization of ribonucleic acid with unique sequences of mouse deoxyribonucleic acid. Biochemistry. 1972 Feb 29;11(5):798–805. doi: 10.1021/bi00755a019. [DOI] [PubMed] [Google Scholar]
- Harris S. E., Schwartz R. J., Tsai M. J., O'Malley B. W., Roy A. K. Effect of estrogen on gene expression in the chick oviduct. In vitro transcription of the ovalbumin gene in chromatin. J Biol Chem. 1976 Jan 25;251(2):524–529. [PubMed] [Google Scholar]
- Harrison P. R., Hell A., Birnie G. D., Paul J. Evidence for single copies of globin genes in the mouse genome. Nature. 1972 Sep 22;239(5369):219–221. doi: 10.1038/239219a0. [DOI] [PubMed] [Google Scholar]
- Honda B. M., Baillie D. L., Candido E. P. The subunit structure of chromatin: characteristics of nucleohistone and nucleoprotamine from developing trout testis. FEBS Lett. 1974 Nov 1;48(1):156–159. doi: 10.1016/0014-5793(74)81086-0. [DOI] [PubMed] [Google Scholar]
- Kedes L. H., Birnstiel M. L. Reiteration and clustering of DNA sequences complementary to histone messenger RNA. Nat New Biol. 1971 Apr 7;230(14):165–169. doi: 10.1038/newbio230165a0. [DOI] [PubMed] [Google Scholar]
- Levenson R. G., Marcu K. B. On the existence of polyadenylated histone mRNA in Xenopus laevis oocytes. Cell. 1976 Oct;9(2):311–322. doi: 10.1016/0092-8674(76)90121-5. [DOI] [PubMed] [Google Scholar]
- Levy B., Johnson C. B., McCarthy B. J. Diversity of sequences in total and polyadenylated nuclear RNA from Drosophila cells. Nucleic Acids Res. 1976 Jul;3(7):1777–1789. doi: 10.1093/nar/3.7.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marzluff W. F., Jr, Huang R. C. Chromatin directed transcription of 5S and tRNA genes. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1082–1086. doi: 10.1073/pnas.72.3.1082. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Milcarek C., Price R., Penman S. The metabolism of a poly(A) minus mRNA fraction in HeLa cells. Cell. 1974 Sep;3(1):1–10. doi: 10.1016/0092-8674(74)90030-0. [DOI] [PubMed] [Google Scholar]
- Nemer M., Graham M., Dubroff L. M. Co-existence of non-histone messenger RNA species lacking and containing polyadenylic acid in sea urchin embryos. J Mol Biol. 1974 Nov 5;89(3):435–454. doi: 10.1016/0022-2836(74)90474-4. [DOI] [PubMed] [Google Scholar]
- Ryffel G. U., McCarthy B. J. Polyadenylated RNA complementary to repetitive DNA in mouse L-cells. Biochemistry. 1975 Apr 8;14(7):1385–1389. doi: 10.1021/bi00678a007. [DOI] [PubMed] [Google Scholar]
- Steggles A. W., Wilson G. N., Kantor J. A., Picciano D. J., Falvey A. K., Anderson W. F. Cell-free transcription of mammalian chromatin: transcription of globin messenger RNA sequences from bone-marrow chromatin with mammalian RNA polymerase. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1219–1223. doi: 10.1073/pnas.71.4.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sullivan D., Palacios R., Stavnezer J., Taylor J. M., Faras A. J., Kiely M. L., Summers N. M., Bishop J. M., Schimke R. T. Synthesis of a deoxyribonucleic acid sequence complementary to ovalbumin messenger ribonucleic acid and quantification of ovalbumin genes. J Biol Chem. 1973 Nov 10;248(21):7530–7539. [PubMed] [Google Scholar]
- Summers J. Physical map of polyoma viral DNA fragments produced by cleavage with a restriction enzyme from Haemophilus aegyptius, endonuclease R-HaeIII. J Virol. 1975 Apr;15(4):946–953. doi: 10.1128/jvi.15.4.946-953.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Suzuki Y., Gage L. P., Brown D. D. The genes for silk fibroin in Bombyx mori. J Mol Biol. 1972 Oct 14;70(3):637–649. doi: 10.1016/0022-2836(72)90563-3. [DOI] [PubMed] [Google Scholar]
- Weinberg E. S., Birnstiel M. L., Purdom I. F., Williamson R. Genes coding for polysomal 9S RNA of sea urchins: conservation and divergence. Nature. 1972 Nov 24;240(5378):225–228. doi: 10.1038/240225a0. [DOI] [PubMed] [Google Scholar]
- Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]
- Zieve G., Penman S. Small RNA species of the HeLa cell: metabolism and subcellular localization. Cell. 1976 May;8(1):19–31. doi: 10.1016/0092-8674(76)90181-1. [DOI] [PubMed] [Google Scholar]