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Cloning and molecular characterization of a telomeric sequence from a temperature-induced Balbiani ring

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Abstract

In Chironomus thummi thummi a Balbiani ring-like structure is induced at the right end of chromosome III by heat shock. Telomere-specific clones were obtained by microcloning this chromosome region. One of the recombinant clones, λCthS, which had an insert of 176 base pairs (bp), hybridized in situ to all the chromosome ends of C. th. thummi except one on the acrocentric chromosome IV. Homologous sequences were also present at two interstitial sites on chromosome II. Southern analysis of genomic DNA indicated that the 176-bp insert represents a basic repeating unit clustered in tandem arrays. Around 1000 copies of this sequence were located in each telomere. The repeats were homogeneous in size but showed some sequence divergence, which seemed to increase when different telomeres were compared. At the nucleotide sequence level the most noticeable feature was the presence of tracts of adenine alternating with tracts of thymine. In situ hybridzation to chromosomal RNA showed that the 176-bp repeat was complementary to the RNA transcribed at the T-BRIII region, and occasionally, to the RNA transcribed at some of the other telomeres during heat shock. Northern blot analysis of total RNA showed that a transcript larger than 10 kilobase pairs (kb) and complementary to the teloeric insert was present in heat-shocked as well as in untreated cells. However, at most only a very weak RNA hybridization was seen at the telomeres of untreated animals.

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References

  • Barettino D, Morcillo G, Díez JL (1982) Induction of heat-shock Balbiani rings after RNA synthesis inhibition in polytene chromosomes of Chironomus thummi. Chromosoma 87:507–517

    Google Scholar 

  • Bauer H (1936) Beiträge zur vergleichenden Morphologie der Speicheldrüsen-Chromosomen (Untersuchungen an den Riesenchromosomen der Dipteren II). Zool Jb Physiol 56:239–276

    Google Scholar 

  • Bedbrook JR, Jones J, O'Dell M, Thompson RD, Flavell RB (1980) A molecular description of telomeric heterochromatin in Secale species. Cell 19:545–560

    Google Scholar 

  • Bergold PJ, Campbell GR, Littau VC, Johnson EM (1983) Sequence and hairpin structure of an inverted repeat series at termini of the Physarum extrachromosomal rDNA molecule. Cell 32:1287–1299

    Google Scholar 

  • Blackburn EH (1984) Telomeres: do the ends justify the means? Cell 37:7–8

    Google Scholar 

  • Blackburn EH, Szostak JW (1984) The molecular structure of centromeres and telomeres. Ann Rev Biochem 53:163–194

    Google Scholar 

  • Blin N, Stafford DW (1976) A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res 3:2302–2308

    Google Scholar 

  • Chan CSM, Tye BK (1983) Organization of DNA sequences and replication origins at yeast telomeres. Cell 33:563–573

    Google Scholar 

  • Clewell DB, Helinski DR (1969) Supercoiled circular DNA protein complex in Escherichia coli: purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166

    Google Scholar 

  • Dulbecco R, Vogt M (1954) Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med 99:167–182

    Google Scholar 

  • Emery HS, Weiner AM (1981) An irregular satellite sequence is found at the termini of the linear extrachromosomal rDNA in Dictyostelium discoideum. Cell 26:411–419

    Google Scholar 

  • Jones JDG, Flavell RB (1983) Chromosomal structure and arrangement of repeated DNA sequences in the telomeric heterochromatin of Secale cereale and its relatives. Cold Spring Harbor Symp Quant Biol 47:1209–1213

    Google Scholar 

  • Kafatos FC, Jones CW, Efstratiadis A (1979) Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res 7:1541–1552

    Google Scholar 

  • Langer-Safer PR, Levine M, Ward DC (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci USA 79:4381–4385

    Google Scholar 

  • Livak KJ, Freund R, Schweber M, Wensink PC, Meselson M (1978) Sequence organization and transcription at two heat-shock loci in Drosophila. Proc Natl Acad Sci USA 75:5613–5617

    Google Scholar 

  • Maniatis T, Hardison RC, Lacy E, Laver J, O'Connell C, Quon D, Sim GK, Efstratiadis A (1978) The isolation of structural genes from libraries of eukaryotic DNA. Cell 15:687–701

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Maxam AH, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–559

    Google Scholar 

  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active DNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acid Res 12:7035–7056

    Google Scholar 

  • Morcillo G, Santa-Cruz MC, Díez JL (1981) Temperature-induced Balbiani rings in Chironomus thummi. Chromosoma 83:341–352

    Google Scholar 

  • Morcillo G, Barettino D, Carretero T, Carmona MJ, Díez JL (1982a) Heat-shock response in Chironomus thummi. Biol Cell 45:298

    Google Scholar 

  • Morcillo G, Barettino D, Díez JL (1982b) Heat sock puffs in isolated salivary glands of Chironomus thummi. Biol Cell 44:221–227

    Google Scholar 

  • Müller F, Walker P, Aeby P, Neuhaus H, Felder H, Back E, Tobler H (1982) Nucleotide sequence of satellite DNA contained in the eliminated genome of Ascaris lumbricoides. Nucleic Acids Res 10:7493–7510

    Google Scholar 

  • Pirrota V, Jäckle H, Edström JE (1983) Genetic engineering: Principles and methods. Setlow JK, Hollaender A (eds). Vol 5. Plenum Publ Corp, New York London, pp 1–17

    Google Scholar 

  • Renkawitz-Pohl R, Bialojan S (1984) A DNA sequence of Drosophila melanogaster with a differential telomeric distribution. Chromosoma 89:206–211

    Google Scholar 

  • Rubin GM (1977) Isolation of a telomeric DNA sequence from Drosophila melanogaster. Cold Spring Harbor Symp Quant Biol 42:1041–1046

    Google Scholar 

  • Rydlander L, Pigon A, Edström JE (1980) Sequences translated by Balbiani ring 75S RNA in vitro present in giant secretory protein from Chironomus tentans. Chromosoma 81:101–113

    Google Scholar 

  • Saiga H, Edström JE (1985) Long tandem arrays of complex repeat units in chironomus telomeres. Embo J (in press)

  • Santa-Cruz MC, Morcillo G, Aller P, Díez JL (1981) Heat-shock induced puffing changes in Chironomus thummi. Cell Differ 10:33–38

    Google Scholar 

  • Scalenghe F, Turco E, Edström JE, Pirrota V, Melli M (1981) Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma 82:205–216

    Google Scholar 

  • Schlesinger MJ, Ashburner M, Tissières A (eds) (1982) Heat-shock: from bacteria to man. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Staller-Young B, Pession A, Traverse KL, French C, Pardue ML (1983) Telomere regions in Drosophila share complex DNA sequences with pericentric heterochromatin. Cell 34:85–94

    Google Scholar 

  • Steinemann M (1984) Telomere repeats within the neo-Y-chromosome of Drosophila miranda. Chromosoma 90:1–5

    Google Scholar 

  • Wobus U (1975) Molecular characterization of an insect genome: Chironomus thummi. Eur J Biochem 59:287–293

    Google Scholar 

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Carmona, M.J., Morcillo, G., Galler, R. et al. Cloning and molecular characterization of a telomeric sequence from a temperature-induced Balbiani ring. Chromosoma 92, 108–115 (1985). https://doi.org/10.1007/BF00328462

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