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Molecular and cytological characterization of repetitive DNA sequences from the centromeric heterochromatin of Sciara coprophila

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Abstract

Sciara coprophila (Diptera, Nematocera) constitutes a classic model to analyze unusual chromosome behavior such as the somatic elimination of paternal X chromosomes, the elimination of the whole paternal, plus non-disjunction of the maternal X chromosome at male meiosis. The molecular organization of the heterochromatin in S. coprophila is mostly unknown except for the ribosomal DNA located in the X chromosome pericentromeric heterochromatin. The characterization of the centromeric regions, thus, is an essential and required step for the establishment of S. coprophila as a model system to study fundamental mechanisms of chromosome segregation. To accomplish such a study, heterochromatic sections of the X chromosome centromeric region from salivary glands polytene chromosomes were microdissected and microcloned. Here, we report the identification and characterization of two tandem repeated DNA sequences from the pericentromeric region of the X chromosome, a pericentromeric RTE element and an AT-rich centromeric satellite. These sequences will be important tools for the cloning of S. coprophila centromeric heterochromatin using libraries of large genomic clones.

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References

  • Abad JP, Carmena M, Baars S, Saunders RDC, Glover DM, Ludeña P, Sentis C, Tyler-Smith C, Villasante A (1992) Dodeca satellite: a conserved G + C-rich satellite from the centromeric heterochromatin of Drosophila melanogaster. Proc Natl Acad Sci U S A 89:4663–4667

    Article  PubMed  CAS  Google Scholar 

  • Abad JP, Agudo M, Molina I, Losada A, Ripoll P, Villasante A (2000) Pericentromeric regions containing 1.688 satellite DNA sequences show anti-kinetochore antibody staining in prometaphase chromosomes of Drosophila melanogaster. Mol Gen Genet 264:371–377

    Article  PubMed  CAS  Google Scholar 

  • Abbott AG, Gerbi SA (1981) Spermatogenesis in Sciara coprophila. II. Precocious chromosome orientation in meiosis II. Chromosoma 83:19–27

    Article  PubMed  CAS  Google Scholar 

  • Abbott AG, Hess JE, Gerbi SA (1981) Spermatogenesis in Sciara coprophila. I. Chromosome orientation on the monopolar spindle of meiosis I. Chromosoma 83:1–18

    Article  PubMed  CAS  Google Scholar 

  • Agudo M, Abad JP, Molina I, Losada A, Ripoll P, Villasante A (2000) A dicentric chromosome of Drosophila melanogaster showing alternate centromere inactivation. Chromosoma 109:190–196

    Article  PubMed  CAS  Google Scholar 

  • Burke WD, Eickbush DG, Xiong Y, Jakubczak J, Eickbush TH (1993) Sequence relationship of retrotransposable elements R1 and R2 within and between divergent insect species. Mol Biol Evol 10:163–185

    PubMed  CAS  Google Scholar 

  • Crouse HV (1943) Translocations in Sciara; their bearing on chromosome behavior and sex determination. Univ Mo Res Bull 379:1–75

    Google Scholar 

  • Crouse HV (1977) X heterochromatin subdivision and cytogenetic analysis in Sciara coprophila (Diptera, Sciaridae) I. Centromere localization. Chromosoma 63:39–55

    Article  Google Scholar 

  • Crouse HV (1979) X heterochromatin subdivision and cytogenetic analysis in S. coprophila (Diptera, Sciaridae). II. The controlling element. Chromosoma 74:219–239

    Article  Google Scholar 

  • Crouse HV, Gerbi SA, Liang CM, Magnus L, Mercer IM (1977) Localization of ribosomal DNA within the proximal X heterochromatin of Sciara coprophila (Diptera, Sciaridae). Chromosoma 64:305–318

    Article  PubMed  CAS  Google Scholar 

  • Gabrusewycz-Garcia N (1964) Cytological and autoradiographic studies in Sciara coprophila salivary gland chromosomes. Chromosoma 15:312–344

    Article  PubMed  CAS  Google Scholar 

  • Gerbi SA (1971) Localization and characterization of rRNA cistrons in Sciara coprophila. J Mol Biol 58:499–511

    Article  PubMed  CAS  Google Scholar 

  • Gerbi SA (1986) Unusual chromosome movements in Sciarid flies. In: Hennig W (ed) Results and problems in cell differentiation, vol 13, Germ line—soma differentiation. Springer, Berlin, pp 71–104

    Google Scholar 

  • Gerbi SA (2007) Helen Crouse (1914–2006): imprinting and chromosome behavior. Genetics 175:1–6

    PubMed  CAS  Google Scholar 

  • Gerbi SA, Crouse HV (1976) Further studies on the ribosomal RNA cistrons of Sciara coprophila (Diptera). Genetics 83:81–90

    PubMed  CAS  Google Scholar 

  • Goday C, Esteban MR (2001) Chromosome elimination in sciarid flies. BioEssays 23:242–250

    Article  PubMed  CAS  Google Scholar 

  • Jordan BR, Latil-Damotte M, Jourdan R (1980) Coding and spacer sequences in the 5.8S-2S region of Sciara coprophila ribosomal DNA. Nucleic Acids Res 8:3565–3573

    Article  PubMed  CAS  Google Scholar 

  • Kerrebrock AW, Srivastava R, Gerbi SA (1989) Isolation and characterization of ribosomal DNA variants from Sciara coprophila. J Mol Biol 210:1–13

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Tamura K, Jakobsen IB, Nei M (2001) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17:1244–1245

    Article  PubMed  CAS  Google Scholar 

  • Lohe AR, Hilliker AJ, Roberts PA (1993) Mapping simple repeated DNA sequences in the heterochromatin of Drosophila melanogaster. Genetics 134:1149–1174

    PubMed  CAS  Google Scholar 

  • Losada A, Abad JP, Villasante A (1997) Organization of DNA sequences near the centromere of the Drosophila melanogaster Y chromosome. Chromosoma 106:503–512

    Article  PubMed  CAS  Google Scholar 

  • Malik HS, Eickbush TH (1998) The RTE class of non-LTR retrotransposons is widely distributed in animals and is the origin of many SINEs. Mol Biol Evol 15:1123–1134

    PubMed  CAS  Google Scholar 

  • Malik HS, Burke WD, Eickbush TH (1999) The age and evolution of non-LTR retrotransposable elements. Mol Biol Evol 16:793–805

    PubMed  CAS  Google Scholar 

  • Méndez-Lago M, Wild J, Whitehead SL, Tracey A, de Pablos B, Rogers J, Szybalski W, Villasante A (2009) Novel sequencing strategy for repetitive DNA in a Drosophila BAC clone reveals that the centromeric region of the Y chromosome evolved from a telomere. Nucleic Acids Res 37:2264–2273

    Article  PubMed  Google Scholar 

  • Metz CW (1925) Chromosome behavior in Sciara (Diptera). Anat Rec 31:346–347

    Google Scholar 

  • Metz CW (1926) Genetic evidence of a selective segregation of chromosomes in Sciara (Diptera). Proc Natl Acad Sci U S A 12:690–692

    Article  PubMed  CAS  Google Scholar 

  • Metz CW (1938) Chromosome behavior, inheritance, and sex determination in Sciara. Am Nat 72:485–520

    Article  Google Scholar 

  • Nagaki K, Cheng Z, Ouyang S, Talbert PB, Kim M, Jones KM, Henikoff S, Buell CR, Jiang J (2004) Sequencing of a rice centromere uncovers active genes. Nat Genet 36:138–145

    Article  PubMed  CAS  Google Scholar 

  • Renkawitz R, Gerbi SA, Glätzer KH (1979) Ribosomal DNA of fly Sciara coprophila has a very small and homogeneous repeat unit. Mol Gen Genet 173:1–13

    Article  PubMed  CAS  Google Scholar 

  • Renkawitz-Pohl R, Matsumoto L, Gerbi SA (1981) Two distinct intervening sequences in different ribosomal DNA repeat units of Sciara coprophila. Nucleic Acids Res 9:3747–3764

    Article  PubMed  CAS  Google Scholar 

  • Rieffel SM, Crouse HV (1966) The elimination and differentiation of chromosomes in the germ line of Sciara. Chromosoma 19:231–276

    Article  PubMed  CAS  Google Scholar 

  • Rovira C, Beermann W, Edström JE (1993) A repetitive DNA sequence associated with the centromeres of Chironomus pallidivittatus. Nucleic Acids Res 21:1775–1781

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Schueler MG, Higgins AW, Rudd MK, Gustashaw K, Willard HF (2001) Genomic and genetic definition of a functional human centromere. Science 294:109–115

    Article  PubMed  CAS  Google Scholar 

  • Serna E, Gorab E, Ruiz MF, Goday C, Eirin-Lopez JM, Sanchez L (2004) The gene sex-lethal of the Sciaridae family (order Diptera, suborder Nematocera) and its phylogeny in dipteran insects. Genetics 168:907–921

    Article  PubMed  CAS  Google Scholar 

  • Sun X, Le HD, Wahlstrom JM, Karpen GH (2003) Sequence analysis of a functional Drosophila centromere. Genome Res 13:182–194

    Article  PubMed  CAS  Google Scholar 

  • Telenius H, Carter NP, Bebb CE, Nordenskjold M, Ponder BA, Tunnacliffe A (1992) Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer. Genomics 13:718–725

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

    Article  PubMed  CAS  Google Scholar 

  • Ware VC, Renkawitz R, Gerbi SA (1985) rRNA processing: removal of only nineteen bases at the gap between 28S alpha and 28S beta rRNAs in Sciara coprophila. Nucleic Acids Res 13:3581–3597

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants BFU2008-02947-C02-02/BMC to C.G. and BFU2008-02947-C02-01/BMC to A.V. from the Ministerio de Ciencia e Innovación, an institutional grant from the Fundación Ramón Areces to the Centro de Biología Molecular “Severo Ochoa” and a grant supporting the Cambridge Resource Centre from the Wellcome Trust to M.A.FS; M.C.Escribá was supported by the FPI fellowship BES-2006-12492 from the Ministerio de Ciencia e Innovación.

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Correspondence to Clara Goday or Alfredo Villasante.

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Communicated by C. Lehner

The GenBank accession numbers for the sequences reported in this paper are HQ883494-513.

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Escribá, M.C., Greciano, P.G., Méndez-Lago, M. et al. Molecular and cytological characterization of repetitive DNA sequences from the centromeric heterochromatin of Sciara coprophila . Chromosoma 120, 387–397 (2011). https://doi.org/10.1007/s00412-011-0320-2

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