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Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma

IV. Heterogeneity and complexity of the nuclear genome

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Abstract

DNA isolated from purified nuclei of Polytoma obtusum has a buoyant density of 1.711 g/ml in CsCl, a Tm of 91.3° C in SSC, and a G + C content of 52.5% as determined by base composition analysis. Thermal dissociation and reassociation studies indicated that this nuclear DNA contains a considerable amount of heterogeneity. Under appropriate reannealing conditions for denatured DNA, about 15% of the DNA reannealed to form a satellite peak at a density of 1.711 g/ml within one hour. Native DNA fractions of different average buoyant densities, ranging from 1.723 to 1.708 g/ml were also obtained in a preparative CsCl gradient, indicating the presence of intermolecular heterogeneity at a molecular size of 8.5×106 daltons. The nuclear DNA reassociated as three distinct classes. The very fast species constituted about 20 % of the total hyperchromicity, the class of intermediate rate comprised roughly 10% of the nuclear DNA, while the remaining 70% consisted of unique sequences. The haploid genome set was estimated by renaturation kinetics studies to contain 5.0×1010 daltons of DNA or 7.5×107 nucleotide pairs. The analytical complexity of the total nuclear genome was found to be 9.35×1010 daltons, thus indicating that vegetative cells of P. obtusum are diploid.

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References

  • Bastia, D.: Studies on the structure of the chloroplast, the nucleic acids of the chloroplast, and the nucleus of Chlamydomonas reinhardtii. Ph.D. Dissertation, University of Chicago, Chicago 1971

    Google Scholar 

  • Bonner, T. I., Brenner, D. J., Neufeld, B. R., Britten, R. J.: Reduction in the rate of DNA reassociation by sequence divergence. J. molec. Biol. 81, 123–136 (1973)

    Google Scholar 

  • Bostock, C.: Repetitious DNA. Advanc. Cell Biol. 2, 153–221 (1971)

    Google Scholar 

  • Britten, R., Kohne, D.: Repeated nucleotide sequences. Carnegie Inst. Wash. Yearbook 66, 68–95 (1967)

    Google Scholar 

  • Britten, R., Kohne, D.: Repeated sequences in DNA. Science 161, 529–540 (1968)

    Google Scholar 

  • Burton, K.: A study of the conditions and mechanisms of the diphenylamine reaction for the colorimetric estimation of DNA. Biochem. J. 62, 315–323 (1956)

    Google Scholar 

  • Chargaff, E., Davidson, J. N.: The nucleic acids, vol. 1, p. 359. New York: Academic Press 1955a

    Google Scholar 

  • Chargaff, E., Davidson, J. N.: The nucleic acids, vol.2, p. 194. New York: Academic Press 1955b

    Google Scholar 

  • Chiang, K.-S., Sueoka, N.: Replication of chloroplast DNA in Chlamydomonas reinhardtii during vegetative cell cycle: Its mode and regulation. Proc. nat. Acad. Sci. (Wash.) 57, 1506–1513 (1967a)

    Google Scholar 

  • Chiang, K.-S., Sueoka, N.: Replication of chromosomal and cytoplasmic DNA during mitosis and meiosis in the eukaryote Chlamydomonas reinhardtii. J. Cell Physiol. 70, Suppl. 1, 89–112 (1967b)

    Google Scholar 

  • Church, R. B., Georgiev, G. P.: Double-stranded regions in renatured DNA from mouse cells. Molec. Biol. Reports 1, 21–25 (1973)

    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. molec. Biol. 77, 1–24 (1973)

    Google Scholar 

  • Gall, J., Cohen, E. H., Polan, M. L.: Repetitive DNA sequences in Drosophila. Chromosoma (Berl.) 33, 319–344 (1971)

    Google Scholar 

  • Gillis, M., DeLey, J., DeCleene, M.: The determination of molecular weight of bacterial genome DNA from renaturation rates. Europ. J. Biochem. 12, 143–153 (1970)

    Google Scholar 

  • Hennig, W., Walker, P. M. B.: Variations in the DNA from two rodents families. Nature (Lond.) 225, 915–919 (1970)

    Google Scholar 

  • Jelinek, W., Darnell, J. E.: Double-stranded regions in heterogeneous nuclear RNA from HeLa cells. Proc. nat. Acad. Sci. (Wash.) 69, 2537–2541 (1972)

    Google Scholar 

  • Kieras, F., Chiang, K.-S.: Characterization of DNA components from some colorless algae. Exp. Cell Res. 64, 89–96 (1971)

    Google Scholar 

  • Kohne, D., Britten, R. J.: Hydroxyapatite techniques for nucleic acid reassociation. In: Procedures in nucleic acid research (Cantoni, G. L., Davies, D. R. eds.), p. 500–512. New York: Academic Press 1971

    Google Scholar 

  • Mandel, M., Marmur, J.: Use of ultraviolet absorbance-temperature profile for determining the guanine plus cytosine content of DNA. In: Methods in enzymology (Grossman, L., Moldave, K., eds.), vol. 12B, 184–195. New York: Academic Press 1968

    Google Scholar 

  • Musich, P.: The distribution of rapidly renaturing DNA in the genome of Oncopeltus. Ph.D. Dissertation, University of Chicago, Chicago 1973

    Google Scholar 

  • Rae, P. M. M.: Chromosomal distribution of rapidly reannealing DNA in Drosophila melanogaster. Proc. nat. Acad. Sci. (Wash.) 67, 1018–1025 (1970)

    Google Scholar 

  • Rae, P. M. M.: The distribution of repetitive DNA sequences in chromosomes. Advanc. Cell and molec. Biol. 2, 74–149 (1972)

    Google Scholar 

  • Ryskov, A. P., Saunders, G. F., Farashyan, V. F., Georgiev, G. P.: Double helical regions in nuclear precursor of mRNA. Biochim. biophys. Acta (Amst.) 312, 152–164 (1973)

    Google Scholar 

  • Schildkraut, C. L., Maio, J. J.: Fractions of HeLa DNA differing in their content of guanine and cytosine. J. molec. Biol. 46, 305–312 (1969)

    Google Scholar 

  • Sinclair, J., Brown, D. D.: Retention of common nucleotide sequences in the ribosomal deoxyribonucleic acid of eukaryotes and some of their physical characteristics. Biochemistry (Wash.) 10, 2761–2769 (1971)

    Google Scholar 

  • Siu, C.-H., Chiang, K.-S., Swift, H.: Novel properties of a dAT-rich DNA satellite in Polytoma obtusum. J. Cell Biol. 55, 241a (1972)

  • Siu, C.-H., Chiang, K.-S., Swift, H.: Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma. III. Leucoplast and nuclear ribosomal RNA cistones. J. Cell Biol. (submitted 1974a)

  • Siu, C.-H., Swift, H., Chiang, K.-S.: Characterization of cytoplasmic and nuclear genomes of the colorless alga Polytoma. I. Ultrastructural analysis of organelles. J. Cell Biol. (submitted 1974b)

  • Siu, C.-H., Swift, H., Chiang, K.-S.: Characterization of cytoplasmic and nuclear genomes of the colorless alga Polytoma. II. Organelle fractionation and properties of their nucleic acids. J. Cell Biol. (submitted 1974c)

  • Studier, F. W.: Sedimentation studies of the size and shape of DNA. J. molec. Biol. 11, 373–390 (1965)

    Google Scholar 

  • Sueoka, N., Chiang, K.-S., Kates, J. R.: DNA replication in meiosis of Chlamydomonas reinhardtii. I. Isotopic transfer experiments with a strain producing light zoospores. J. molec. Biol. 25, 47–66 (1967)

    Google Scholar 

  • Suzuki, Y., Gage, L. P., Brown, D. D.: The genes of silk fibroin in Bombyx mori. J. molec. Biol. 70, 637–649 (1972)

    Google Scholar 

  • Walker, P. M. B.: How different are the DNAs from related animals? Nature (Lond.) 219, 228 (1968)

    Google Scholar 

  • Wellauer, P. K., Dawid, I. B.: Secondary maps of RNA: Processing of HeLa ribosomal RNA. Proc. nat. Acad. Sci. (Wash.) 70, 2827–2831 (1973)

    Google Scholar 

  • Wells, R., Sager, R.: Denaturation and the renaturation kinetics of chloroplast DNA from Chlamydomonas reinhardtii. J. molec. Biol. 58, 611–622 (1971)

    Google Scholar 

  • Wetmur, J. G., Davidson, N.: Kinetics of renaturation of DNA. J. molec. Biol. 31, 349–370 (1968)

    Google Scholar 

  • Wilson, D. A., Thomas, C. A.: Palindromes in chromosomes. J. molec. Biol. 84, 115–144 (1974)

    Google Scholar 

  • Wu, J.-R., Hurn, J., Bonner, J.: Size and distribution of the repetitive segments of the Drosophila genome. J. molec. Biol. 64, 211–219 (1972)

    Google Scholar 

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Siu, CH., Chiang, KS. & Swift, H. Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma . Chromosoma 48, 19–40 (1974). https://doi.org/10.1007/BF00284864

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