Abstract
Intermediate repetitive sequences of Plethodon cinereus which comprised about 30% of the genomic DNA were isolated and iodinated with 125I. About 5% of the 125I-repetitive fraction hybridized with a large excess of DNA from P. dunni at Cot 20. About half of the 125I-DNA in the hybrids was resistant to extensive digestion with S-1 nuclease. The average molecular size of the S-1 nuclease-resistant fraction was about 100 nucleotide pairs. The melting temperature of the S-1 nuclease-resistant fraction was about 2° lower than that of the corresponding fraction made with P. cinereus DNA. These results are taken to indicate the presence in the genomes of P. cinereus and P. dunni of evolutionarily stable “common” repetitive sequences. The average frequency of repetition of the common repetitive sequences is about 6,000 × in both species. The common repetitive fraction is also present in the genomes of other species of Plethodon, although the general populations of intermediate repetitive sequences are markedly different from one species to another. The cinereus-dunni common repetitive sequences could not be detected in plethodontids belonging to different tribes, nor in more distantly related amphibians. The profiles of binding of the common repetitive sequences to CsCl or Cs2SO4-Ag+ density gradient fractions of P. dunni DNA suggested that these sequences consisted of heterogeneous components with respect to base compositions, and that they did not include large amounts of the genes for ribosomal RNA, 5S RNA, 4S RNA, or histone messenger RNA. — In situ hybridization of the 3H-labelled intermediate repetitive sequences of P. cinereus to male meiotic chromosomes of the same species gave autoradiographs after an exposure of seven days showing all 14 chromosomes labelled. The pattern of labelling appeared not to be random, but was impossible to analyse on account of the irregular shapes and different degrees of stretching of diplotene and prometaphase chromosomes. In situ hybridization of the same sequences to meiotic chromosomes from P. dunni gave autoradiographs after 60 d exposure in which all chromosomes were labelled. These heterologous in situ hybrids can only have involved the “common” repetitive sequences.
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References
Ando, T.: A nuclease specific for heat-denatured DNA isolated from a product of Aspergillus oryzae. Biochim. biophys. Acta (Amst.) 114, 158–168 (1966)
Britten, R.J., Davidson, E.H.: Gene regulation for higher cells: A theory. Science 165, 349–357 (1969)
Brown, D.D., Weber, C.S.: Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4S RNA, 5S RNA and ribosomal RNA. J. molec. Biol. 34, 661–680 (1968)
Brown, D.D., Wensink, P.C., Jordan, E.: Purification and some characteristics of 5S DNA from Xenopus laevis. Proc. nat. Acad. Sci. (Wash.) 68, 3175–3179 (1971)
Church, R.B., Schulz, G.A.: Differential gene activity in the pre- and post-implantation mammalian embryo. In: Current topics in developmental biology (A.A. Moscona and A. Monroy, eds.), Vol. 8, pp. 179–202. New York: Academic Press 1974
Commerford, S.L.: Iodination of nucleic acids in vitro. Biochemistry 10, 1993–1999 (1971)
Davidson, E.H., Britten, R.J.: Organization, transcription, and regulation in the animal genome. Quart. Rev. Biol. 48, 565–613 (1973)
Davidson, E.H., Galau, G.A., Angerer, R.C., Britten, R.J.: Comparative aspects of DNA organization in metazoa. Chromosoma (Berl.) 51, 253–259 (1975a)
Davidson, E.H., Hough, B.R., Klein, W.H., Britten, R.J.: Structural genes adjacent to interspersed repetitive DNA sequences. Cell 4, 217–238 (1975b)
Denhardt, D.T.: A membrane-filter technique for the detection of complementary DNA. Biochem biophys. Res. Commun. 23, 641–646 (1966)
Farquhar, M.N., McCarthy, B.J.: Evolutionary stability of the histone genes of sea urchins. Biochemistry 12, 4113–4122 (1973)
Gilbert, W., Maizels, N., Maxam, A.: Sequences of controlling regions of the lactose operon. Cold. Spr. Harb. Symp. quant. Biol. 38, 845–855 (1973)
Goldberg, R.B., Crain, W.R., Ruderman, J.V., Moore, G.P., Barnett, T.R., Higgins, R.C., Gelfand, R.A., Galau, G.A., Britten, R.J., Davidson, E.H.: DNA sequence organization in the genomes of five marine invertebrates. Chromosoma (Berl.) 51, 225–251 (1975)
Graham, D.E., Neufeld, B.R., Davidson, E.H., Britten, R.J.: Interspersion of repetitive and non-repetitive DNA sequences in the sea urchin genome. Cell 1, 127–137 (1974)
Hennen, S., Mizuno, S., Macgregor, H.C.: In situ hybridization of ribosomal DNA labelled with 125Iodine to metaphase and lampbrush chromosomes from newts. Chromosoma (Berl.) 50, 349–369 (1975)
Holmes, D.S., Bonner, J.: Interspersion of repetitive and single-copy sequences in nuclear ribonucleic acid of higher molecular weight. Proc. nat. Acad. Sci. (Wash.) 71, 1108–1112 (1974)
Kedes, L.H., Birnstiel, M.L.: Reiteration and clustering of DNA sequences complementary to histone messenger RNA. Nature (Lond.) New Biol. 230, 165–169 (1971)
Lagowski, J.M., Yu, M.-Y.W., Forrest, H.S., Laird, C.D.: Dispersity of repeat DNA sequences in Oncopeltus fasciatus, an organism with diffuse centromeres. Chromosoma (Berl.) 43, 349–373 (1973)
Macgregor, H.C., Kezer, J.: The chromosomal localization of a heavy satellite DNA in the testis of Plethodon c. cinereus. Chromosoma (Berl.) 33, 167–182 (1971)
Macgregor, H.C., Mizuno, S.: In situ hybridization of “nick-translated” 3H-ribosomal DNA to chromosomes from salamanders. Chromosoma (Berl.) 54, 15–25 (1976)
Maniatis, T., Ptashne, M.: Multiple repressor binding at the operators in bacteriophage λ. Proc. nat. Acad. Sci. (Wash.) 70, 1531–1535 (1973)
Manning, J.E., Schmid, C.W., Davidson, N.: Interspersion of repetitive and nonrepetitive DNA sequences in the Drosophila melanogaster genome. Cell 4, 141–155 (1975)
Mizuno, S., Macgregor, H.C.: Chromosomes, DNA sequences, and evolution in salamanders of the genus Plethodon. Chromosoma (Berl.) 48, 239–296 (1974)
Smith, M.J., Hough, B.R., Chamberlin, M.E., Davidson, E.H.: Repetitive and non-repetitive sequences in sea urchin heterogeneous nuclear RNA. J. molec. Biol. 85, 103–126 (1974)
Staynov, D.Z., Pinder, J.C., Gratzer, W.B.: Molecular weight determination of nucleic acids by gel electrophoresis in non-aqueous solution. Nature (Lond.) New Biol. 235, 108–110 (1972)
Studier, F.W.: Sedimentation studies of the size and shape of DNA. J. molec. Biol. 11, 373–390 (1965)
Ullman, J.S., McCarthy, B.J.: The relationship between mismatched base pairs and the thermal stability of DNA duplexes. II. Effects of deamination of cytosine. Biochim. biophys. Acta (Amst.) 294, 416–424 (1973)
Wang, C.H., Jones, D.E.: Liquid scintillation counting of paper chromatograms. Biochem. biophys. Res. Commun. 1, 203–205 (1959)
Weinberg, E.S., Birnstiel, M.L., Purdom, I.F., Williamson, R.: Genes coding for polysomal 9S RNA of sea urchins: Conservation and divergence. Nature (Lond.) 240, 225–228 (1972)
Weinberg, R.A., Loening, U., Willems, M., Penman, S.: Acrylamide gel electrophoresis of HeLa cell nucleolar RNA. Proc. nat. Acad. Sci. (Wash.) 58, 1088–1095 (1967)
Wensink, P.C., Finnegan, D.J., Donelson, J.E., Hogness, D.S.: A system for mapping DNA sequences in the chromosomes of Drosophila melanogaster. Cell 3, 315–325 (1974)
Whiteley, H.R., McCarthy, B.J., Whiteley, A.H.: Conservatism of base sequences in RNA for early development of echinoderms. Develop. Biol. 21, 216–242 (1970)
Whiteley, H.R., Mizuno, S., Lee, Y.R., Whiteley, A.H.: Transcripts of reiterated DNA sequences in the determination of blastomeres and early differentiation in echinoid larvae. Amer. Zool. 15, 619–648 (1975)
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Mizuno, S., Andrews, C. & Macgregor, H.C. Interspecific “common” repetitive DNA sequences in salamanders of the genus Plethodon . Chromosoma 58, 1–31 (1976). https://doi.org/10.1007/BF00293437
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DOI: https://doi.org/10.1007/BF00293437