Abstract
Replication rate and replicon sizes in chromosomal DNA of in vitro cultured diploid D. melanogaster cells were determined using autoradiography of 3H-thymidine labeled DNA. Synthesis of DNA in euchromatic and heterochromatic regions of Drosophila diploid cells occurs at different periods of the S phase which lasts 10 h. During the first 4 h the synthesis is observed only in euchromatic regions. The heterochromatic synthesis starts shortly before the synthesis in euchromatic regions is completed and lasts for 6 h until the end of the S phase. The cells were synchronized by 5fluorodeoxyuridine which blocked the diploid cell DNA synthesis. Synthesis was found to start simultaneously in most euchromatic replicons. In the majority of the replicons the synthesis started at a single point and proceeded bidirectionally. The average rate of DNA synthesis per fork was 12.5 μm/h (38 kb). The mean distance between the middle points of adjacent labeled regions was 70 μm (210 kb). The size of most replicons ranged from 40 to 120 μm. — These estimates do not apply to the heterochromatic portions of the D. melanogaster genome since the measurements have been carried out on DNA preparations obtained during the first 2 h of the S phase. — On the average, a replicon can consist of 7 chromomeres since the size of a replicon in diploid cell chromosomal DNA and DNA length of a polytene chromomere average 210 and 30 kb, respectively.
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Amaldi, F., Carnevali, G., Leoni, L., Marioti, D.: Replicon origines in Chinese hamster cell DNA. I. Labeling procedure and preliminary observation. Exp. Cell Res. 74, 367–374 (1972)
Ananiev, E.V., Gvozdev, V.A.: Differences in DNA replication in the X-chromosome of male, female and intersexes of Drosophila melanogaster. Chromosoma (Berl.) 49, 233–241 (1975)
Barigozzi, C., Dolfini, S., Fraccaro, M., Rezzonico Raimondi, G., Tiepolo, L.: In vitro study of the DNA replication patterns of somatic chromosomes of Drosophila melanogaster. Exp. Cell Res. 43, 231 (1966)
Beermann, W.: Structure and function of interfase chromosomes. Genetics today 2, 373–384 (1965)
Blumenthal, A.B., Kriegstein, H.J., Hogness, D.S.: The units of DNA replication in Drosophila melanogaster chromosomes. Cold Spr. Harb. Symp. quant. Biol. 38, 205–224 (1938)
Bridges, C.B.: Revised salivary gland map of X-chromosomes of Drosophila melanogaster. J. Hered. 29, 11–13 (1938)
Gallan, H.G.: Replication of DNA in the chromosomes of eukaryotes. Proc. roy. Soc. (London) 181, 19–41 (1972)
Cairns, J.: Autoradiography of Hela cell DNA. J. molec. Biol. 15, 372–373 (1966)
Chan, A.C., Walker, O.G.: DNA synthesis in L-cells in the presence of 5-fluorodeoxyuridine. Biochem. biophys. Abst. 395, 422–432 (1975)
Comings, D.E.: Quantitative autoradiography of heterochromatin replication in Microtus agrestis. Chromosoma (Berl.) 29, 434–445 (1970)
Gvozdev, V.A., Gostimsky, S.A., Gerasimova, T.J., Dubrovskaya, Ya.S., Braslavskaya, O.Yu.: Fine structure analysis at the 2D3-2F5 region of the X-chromosome of Drosophila melanogaster. Genetica (Russ.) 10, 73–79 (1975)
Gvozdev, V.A., Kakpakov, V.T.: Culture of embryonic cells of Drosophila melanogaster in vitro. Genetica (Russ.) 2, 129 (1968)
Hand, L., Tamm, J.: Initiation of DNA replication in mammalian cells and its inhibition by reovirus infection. J. molec. Biol. 82, 175–183 (1974)
Hochmann, B.: Analysis of chromosome 4 in Drosophila melanogaster. II. Ethyl methanesulfonate induced lethals. Genetics 67, 235–252 (1971)
Howard, A., Pelc, S.R.: Nuclear incorporation of P32 as demonstrated by autoradiographs. Exp. Cell Res. 2, 173–187 (1951).
Howard, E.F., Plaut, W.: Chromosomal DNA synthesis in Drosophila melanogaster. J. Cell Biol. 39, 415–429 (1968)
Huberman, J.A., Rigs, A.D.: On the mechanism of DNA replication in mammalian chromosomes. J. molec. Biol. 32, 327–341 (1968)
Huberman, J.A., Tsai, A.: Direction of replication in mammalian cells. J. molec. Biol. 75, 5–12 (1973)
Judd, B.M., Shen, M.W., Kaufman, T.C.: The anatomy and function of a segment of the X-Chromosome of Drosophila melanogaster. Genetics 71, 139–156 (1972)
Kakpakov, V.T., Gvozdev, V.A., Platova, T.P., Polukarova, L.G.: In vitro establishment of embryonic cell lines of Drosophila melanogaster. Genetica (Russ.) 5, 67–75 (1969)
Lakhotia, S.C.: Chromosomal basis of dosage compensation in Drosophila melanogaster. II. The DNA replication in an autosome-X insertion in Drosophila melanogaster. Genet. Res. (Camb.) 15, 301–307 (1970)
Lark, K.G., Consigli, R., Toliver, A.: DNA replication in Chinese Hamster cell: evidence for a single replication fork per replicon. J. molec. Biol. 58, 873–875 (1971)
Lifschytz, E., Falk, R.: Fine structure analysis of a chromosome segment in Drosophila melanogaster. Analysis of EMS-induced mutant. Mutation Res. 8, 147–155 (1969)
Lima-de-Faria, A.: Differential uptake of tritiated thymidine into hetero- and euchromatin in Melanopus and Secale. J. biophys. biochem. Cytol. 6, 457–466 (1968)
Lomakina, L.Ya., Kustova, O.V.: Evidence of DNA synthesis in the presence of FUDR and amethophterin. Dokl. Acad. Nauk SSSR, Biol. 181, 463–467 (1968)
Macfarlane, P.W., Callan, H.G.: DNA replication in the chromosomes of the chicken, Gallus domesticus. J. Cell Sci. 13, 821–839 (1973)
Mulder, M.P., Duijn, D. van, Gloor, M.J.: The replication organization of DNA in polytene chromosomes of Drosophila melanogaster hydei. Genetica ('s-Gravenhage) 39, 385–428 (1968)
Rudkin, G.T.: Cytochemistry in the ultraviolet. Microchem. J. Symp., Ser. 1, 261–278 (1961)
Wolstenholme, D.R.: Replicating DNA molecules from eggs of Drosophila melanogaster. Chromosoma (Berl.) 43, 1–18 (1973)
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Ananiev, E.V., Polukarova, L.G. & Yurov, Y.B. Replication of chromosomal DNA in diploid Drosophila melanogaster cells cultured in vitro. Chromosoma 59, 259–272 (1977). https://doi.org/10.1007/BF00292782
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DOI: https://doi.org/10.1007/BF00292782