Abstract
Ever since the method of molecular hybridization of nucleic acids was conceived, a number of biologists have tried to hybridize labeled RNA or DNA to chromosomes for detection by autoradiography. The first report was by French and Kitzmiller (1967), who hybridized 3H-DNA from Drosophila melanogaster to the DNA of the salivary chromosomes of the same species in situ. Although their autoradiographs showed radioactivity on the chromosomes well above background, inconsistent results prevented them from publishing a full-length paper. In a second study by Gall and Pardue (1969) there were more complete and convincing findings using the oöcytes of Xenopus. In this case, tritium labeled rRNA was found to bind to the rDNA of the nucleolus and to the extrachromosomal rDNA of free nucleoli. The hybridization methods were essentially similar to the nitrocellulose filter method of Gillespie and Spiegelman (1965). The labeled hybrid was unaffected by ribonuclease and unlabeled rRNA competed effectively against labeled rRNA for binding sites. A number of advances have been made in the last two years. We will report some new results, principally on hybridization to chromosomes of the various classes of 3H-RNA, and discuss the prospects of the cytological technique for the future. No attempt is made to review the literature but rather we will evaluate the methods as they can be applied to new and challenging experiments.
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References
Ashburner, M.: Patterns of puffing activity in the salivary gland chromosomes of Drosophila. II. The X-chromosome puffing patterns of D. melanogaster and D. simulans. Chromosoma (Berl.) 27, 47–63 (1969a). IV. Variability of puffing patterns. Chromo-soma (Berl.) 27, 156–177 (1969b).
Britten, R.M., Kohne, D.E.: Nucleotide sequence repetition. Carnegie Inst. Wash. Year Book 65, 87–106 (1966).
Britten, R. J., Kohne, D.E.: Repeated sequences in DNA. Science 161, 529–540 (1968).
Brown,D.D., Weber, C. S.: Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4 S, 5 S RNA and ribosomal RNA. J. molec. Biol. 34, 661–680 (1968).
Buongiorno-Nardelli,M., Amaldi, F.: Autoradiographic detection of molecular hybrids between rRNA and DNA in tissue sections. Nature (Lond.) 225, 946–948 (1970).
Cleaver, J.E.: Thymidine metabolism and cell kinetics. In: Neuberger, A., Tatum, E. L. (Eds.): Frontiers of Biology Vol. 6, Amsterdam: North-Holland Publ. Co. 1967.
Darnell, J. E., Pagoulatos, G. N., Lindberg, U., Balint, R.: Studies on the relationship of mRNA to heterogenous nuclear RNA in mammalian cells. Cold. Spr. Harb. Symp. quant. Biol. 35, 555–560 (1970).
Dick, C., Johns, E. W.: The removal of histones from calf thymus deoxyribonucleoprotein and calf thymus tissue with acetic acid containing fixatives. Biochem. J. 105, 46 P (1967).
Eckardt, R. A.: Studies on the chromosomal distribution of repetitive nucleotide sequences in polytene chromosomes. J. Cell Biol. 47, 55a (1970).
French, W. L., Kitzmiller, J. B.: The formation of molecular hybrids in situ on chromosomes of Drosophila melanogaster. Amer. Zoologist 7, 782 (1967).
Gall, J. G.: The genes for ribosomal RNA during oögenesis. Genetics 61, 121–132, Suppl. 1 (1969).
Gall, J. G. Mary Lou Pardue: The formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc. nat. Acad. Sci. (Wash.) 63, 378–383 (1969).
Gall, J. G. Mary Lou Pardue: Nucleic acid hybridization in cytological preparations. Methods in Enzymology 12 C. (in press) (1971).
Gillam, I., Millward, S., Blew, D., von Tigerstrom, M., Wimmer, E., Tener, G. M.: The separation of soluble ribonucleic acids on benzoylated Diethylaminoethylcellulose. Biochemistry 6, 3043–3056 (1967).
Gillespie,D., Spiegelman, S.: A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. Mol. Biol. 12, 829–842 (1965).
Hnilica, L. S., Kappler, H. A., Hnilica, V. S.: Biosynthesis of histones and acidic nuclear proteins under different conditions of growth. Science 150, 1470–1472 (1965).
John, H. A., Birnstiel, M. L., Jones, K. W.: RNA-DNA hybrids at the cytological level. Nature (Lond.) 223, 582–587 (1969).
Jones, K. W.: Chromosomal and nuclear location of mouse satellite DNA in individual cells. Nature (Lond.) 225, 912–915 (1970).
Kaufmann, B. P., Iddles, Marcia K.: Ectopic pairing in Salivary-gland chromosomes of Drosophila. Port. Acta Biol. 7, 225–248 (1965).
Laird, C.D., Mccarthy, B. J.: Magnitude of interspecific nucleotide sequence variability in Drosophila. Genetics 60, 303–322 (1968a).
Laird, C.D., Mccarthy, B. J.: Nucleotide sequence homology within the genome of Drosophila melanogaster. Genetics 60, 323–324 (1968 b).
Lindsley, Danl., Grell, E.H.: Genetic Variations of Drosophila melanogaster. Carnegie Inst. Wash. Publ. No. 627 (1967).
Macgregor, H. C., Kezer, J.: The chromosomal localization of a heavy satellite DNA in the testis of Plethodon c. cinereus. Chromosoma 33, 167–182 (1971).
Mccarthy, B. J., Church, R.B.: The specificity of molecular hybridization reactions. Ann. Rev. Biochem. 39, 131–150 (1970).
Miller, O.L., Beatty, B. R.: Extrachromosomal nucleolar genes in amphibian oöcytes. Genetics 61, (Suppl. 1) 133–144 (1969).
Pardue, M. L., Gall, J. G.: Chromosomal localization of mouse satellite DNA. Science 168, 1356–1358 (1970).
Pardue, M. L., Gerbi, S. A., Eckardt, R. A., Gall, J. G.: Cytological localization of DNA complementary to ribosomal RNA in polytene chromosomes of Diptera. Chromosoma (Berl.) 29, 268–290 (1970).
Pelling, C.: Ribonukleinsäure-synthese der Riesenchromosomen. Autoradiographische Untersuchungen an Chironomus tentans. Chromosoma (Berl.) 15, 71–122 (1964).
Pelling, C., Puff RNA in polytene chromosomes. Cold Spr. Harb. Symp. quant. Biol. 35, 521–531 (1970).
Raepeter M. M.: Chromosomal distribution of rapidly reannealing DNA in Drosophila melanogaster. Proc. nat. Acad. Sci. (Wash.) 67, 1018–1025 (1970).
Ritossa, F.M., Spiegelman, S.: Localization of DNA complementary to ribosomal RNA in the nucleolus organizer region of Drosophila melanogaster. Proc. nat. Acad. Sci. (Wash.) 53, 737–745 (1965).
Ritossa, F.M., Atwood, K. C., Spiegelman, S.: On the redundancy of DNA complementary to amino acid transfer RNA and its absence from the nucleolar organizer region of Drosophila melanogaster. Genetics 54, 663–676 (1966 a).
Ritossa, F.M., Atwood, K. C., Spiegelman, S A molecular explanation of the bobbed mutants of Drosophila as partial deficiencies of “ribosomal” DNA. Genetics 54, 819–834 (1966b).
Steffensen, Dale M., Wimber, D. E.: Localization of tRNA genes in the salivary chromosomes of Drosophila by RNA:DNA Hybridization. Genetics 69, 163–178 (1971).
Tartof, K. D., Perry, R. P.: The 5S RNA genes of Drosophila melanogaster. J. Mol. Biol. 51, 171–183 (1970).
Wimber, D.E., Steffensen, D. M.: Localization of 5S RNA genes in Drosophila chromosomes by RNA-DNA hybridization. Science 170, 639–641 (1970).
Wolstenholme, D. R.: The distribution of RNA and DNA in salivary gland chromosomes of Chironomus tentans as revealed by fluorescence microscopy. Chromosoma (Berl.) 17, 219–229 (1965).
Yasmineh, W. G., Yunis, J. J.: Satellite in DNA mouse heterochromatin. Biochem. Biophys. Res. Commun. 35, 779–782 (1969).
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Steffensen, D.M., Wimber, D.E. (1972). Hybridization of Nucleic Acids to Chromosomes. In: Ursprung, H. (eds) Nucleic Acid Hybridization in the Study of Cell Differentiation. Results and Problems in Cell Differentiation, vol 3. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-37149-6_5
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DOI: https://doi.org/10.1007/978-3-540-37149-6_5
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