Skip to main content
SpringerLink
Log in

Molecular characterization of the B-2 DNA puff gene of Rhynchosciara americana

  • Research Article
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

We have sequenced a 2.5-kb DNA fragment of the B-2 DNA puff from the sciarid Rhynchosciara americana and have defined its transcription unit. This puff is active during the formation of the communal cocoon, which is important for successful metamorphosis of this species and coincides with the final cycle of polytenization in its salivary glands. The B-2 polypeptide, together with the products of two other previously characterized DNA puffs, seems to be engaged in an interaction that results in a gradual modification and hardening of the cocoon structure. The B-2 messenger is temporally regulated in apparent coordination with the other puff products. The predicted polypeptide has characteristics similar to polypeptides from previously sequenced DNA puff genes, in particular those from the R. americana C-8 gene and the Bradysia hygida C-4 gene. The cloned sequence of the B-2 puff is differentially amplified in the three gland regions examined, achieving its highest amplification level of approximately fourfold (two extra cycles) in the anterior segment of the gland. The C-3 DNA puff sequence was also found to be differentially amplified in the different gland regions. Implications of the widespread presence of DNA amplification as a form of gene regulation in the Sciaridae are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Notes

  1. http://bmerc-www.bu.edu/psa/

  2. http://prodes.toulouse.inra.fr/prodom.html

References

  • Alvarenga CA, Winter CE, Stocker AJ, Pueyo MT, Lara FJ (1991) In vivo effects of ecdysterone on puff formation, and RNA and protein synthesis in the salivary glands of Rhynchosciara americana. Braz J Med Biol Res 24:985–1002

    CAS  PubMed  Google Scholar 

  • Amabis DC, Amabis JM (1984a) Hormonal control of gene amplification in the salivary gland chromosomes of Trichosia pubescens. Dev Biol 102:10–20

    CAS  PubMed  Google Scholar 

  • Amabis DC, Amabis JM (1984b) Effects of ecdysone in polytene chromosomes of Trichosia pubescens. Dev Biol 102:1–9

    CAS  PubMed  Google Scholar 

  • Berendes H, Lara FJS (1975) RNA synthesis: a requirement for hormone-induced DNA amplification in Rhynchosciara americana. Chromosoma 50:259–274

    CAS  PubMed  Google Scholar 

  • Bonaldo MF, Santelli RV, Lara FJS (1979) The transcript from a DNA puff of Rhynchosciara and its migration to the cytoplasm. Cell 17:824–833

    Article  Google Scholar 

  • Breuer ME, Pavan C (1955) Behaviour of polytene chromosomes of Rhynchosciara angelae at different stages of larval development. Chromosoma 7:371–386

    Google Scholar 

  • Calvi BR, Spradling AC (1999) Chorion gene amplification in Drosophila: a model for origin of DNA replication and S-phase control. In: Fischer P (ed) Genetic approaches to eukaryotic replication and repair. Academic, New York, pp 407–417

    Google Scholar 

  • Chen C-YA, Shyu A-B (1995) AU-rich elements: characterization and importance in mRNA degradation. Trends Biochem Sci 20:465–470

    Article  CAS  PubMed  Google Scholar 

  • De Toledo SM, Lara FJS (1978) Translation of messages transcribed from DNA puffs of Rhynchosciara. Biochem Biophys Res Commun 85:160–166

    PubMed  Google Scholar 

  • Dreyfus A, Nonato E, Breuer ME, Pavan C (1951) Cromossomos politênicos em vários órgãos de Rhynchosciara angelae. Rev Bras Biol 11:439–450

    Google Scholar 

  • Ewing B, Green P (1998) Base-calling of automated sequencer traces using Phred. 2. Error probabilities. Genome Res 8:186–194

    CAS  PubMed  Google Scholar 

  • Ewing B, Hillier LD, Wendl MC, Phil Green P (1998) Base-calling of automated sequencer traces using Phred. 1. Accuracy assessment. Genome Res 8:175–185

    CAS  PubMed  Google Scholar 

  • Ficq A, Pavan C (1957) Autoradiography of polytene chromosomes of Rhynchosciara angelae at different stages of larval development. Nature 180:983–984

    CAS  PubMed  Google Scholar 

  • Fontes AM, Conacci ME, Monesi N, deAlmeida JC, Paço-Larson ML (1999) The DNA puff BhB10-1 gene encodes a glycine-rich protein secreted by late stage larval salivary glands of Bradysia hygida. Gene 231:67–75

    Article  CAS  PubMed  Google Scholar 

  • Fresquez CL (1979) Nucleic acid synthesis in Rhynchosciara hollaenderi polytene chromosomes I. Dose response and temporal sequence after injection of 20-hydroxy-ecdysone. Insect Biochem 9:517–523

    Article  CAS  Google Scholar 

  • Frydman HM, Cadavid EO, Yokosawa J, Silva FH, Navarro-Cattapan LD, Santelli RV, Jacobs-Lorena M, Graessmann M, Graessmann A, Stocker AJ, Lara FJS (1993) Molecular characterization of the DNA puff C-8 gene of Rhynchosciara americana. J Mol Biol 233:99–803

    Article  Google Scholar 

  • Furger A, O’Sullivan JM, Binnie A, Lee BA, Proudfoot NJ (2002) Promoter proximal splice sites enhance transcription. Genes Dev 16:2792–2799

    Article  CAS  PubMed  Google Scholar 

  • Gerbi SA, Strezoska Z, Waggener JM (2002) Initiation of DNA replication in multicellular eukaryotes. J Struct Biol 140:17–30

    Article  CAS  PubMed  Google Scholar 

  • Glover DM, Zaha A, Stocker AJ, Santelli RV, Pueyo MT, deToledo SM, Lara FJS (1982) Gene amplification in Rhynchosciara salivary glands chromosomes. Proc Natl Acad Sci USA 79:2947–2951

    CAS  PubMed  Google Scholar 

  • Gordon D, Abajian C, Green P (1998) Consed: a graphical tool for sequence finishing. Genome Res 8:195–202

    CAS  PubMed  Google Scholar 

  • Hultmark D, Klemenz R, Gehring WJ (1986) Translational and transcriptional control elements in the untranslated leader of heat shock gene hsp-22. Cell 44:429–438

    Article  CAS  PubMed  Google Scholar 

  • Lara FJS, Tamaki H, Pavan C (1965) Laboratory culture of Rhynchosciara angelae. Am Nat 99:189–191

    Article  Google Scholar 

  • Liang C, Spitzer JD, Smith HS, Gerbi SA (1993) Replication initiates at a confined region during DNA amplification in Sciara DNA puff II/9A. Genes Dev 7:1072–1084

    CAS  PubMed  Google Scholar 

  • Lobo ES (2000) Pufe de DNA C4B de Trichosia pubecens: caracterização molecular de um amplicon e sequenciamento do gene C4B-2. PhD Thesis Intituto de Biociências-Universidade São Paulo

    Google Scholar 

  • Machado-Santelli GM, Basile R (1975) DNA replication and DNA puffs in salivary chromosomes of Rhynchosciara. Cienc Cult 27:167–174

    CAS  Google Scholar 

  • Millar S, Hayward D, Read CA, Browne MJ, Santelli RV, Garcia-Vallego F, Pueyo MT, Zaha A, Glover DM, Lara FJS (1985) Segments of chromosomal DNA from Rhynchosciara americana that undergo additional rounds of DNA replication in the salivary gland DNA puffs have only weak ARS activity in yeast. Gene 34:81–86

    Article  CAS  PubMed  Google Scholar 

  • Ockretic MC, Penoni JS, Lara FJS (1977) Messenger-like RNA synthesis and DNA chromosomal puffs in the salivary glands of Rhynchosciara americana. Arch Biochem Biophys 178:158–165

    PubMed  Google Scholar 

  • Penalva LO, Yokosawa J, Stocker AJ, Soares MA, Graessmann M, Orlando TC, Winter CE, Botella LM, Graessmann A, Lara FJ (1997) Molecular characterization of the C-3 DNA puff gene of Rhynchosciara americana. Gene 193:163–172

    Article  CAS  PubMed  Google Scholar 

  • Pirrotta V, Bonaldo MF, Amabis JM, Santelli RV, Frydman HM, Cattapan LDN, Lara FJS (1988) Cloning of sequences from the B-2 DNA puff of Rhynchosciara salivary glands chromosomes. Arq Biol Technol (Curitiba) 31:41

    Google Scholar 

  • Rice P, Longden I, Bleasby A (2000) EMBOSS: the European molecular biology open software suite. Trends Genet 16:276–277

    Article  CAS  PubMed  Google Scholar 

  • Rudkin GT, Corlette SL (1957) Disproportionate synthesis of DNA in a polytene chromosome region. Proc Natl Acad Sci USA 43:964–968

    CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) A laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Santelli RV, Machado-Santelli GM, Pueyo MT, Navarro-Cattapan LD, Lara FJ (1991) Replication and transcription in the course of DNA amplification of the C3 and C8 DNA puffs of Rhynchosciara americana. Mech Dev 36:59–66

    Article  CAS  PubMed  Google Scholar 

  • Shaw G, Kamen R (1986) A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 46:659–667

    Article  CAS  PubMed  Google Scholar 

  • Soares MB, Bonaldo MF, Jelenc P, Su L, Lawton L, Efstratiadis A (1994) Construction and characterization of a normalized cDNA library. Proc Natl Acad Sci USA 91:9228–9232

    CAS  PubMed  Google Scholar 

  • Spradling AC (1981) The organization and amplification of two chromosomal domains containing Drosophila chorion genes. Cell 27:193–201

    Article  CAS  PubMed  Google Scholar 

  • Spradling AC, Mahowald AP (1980) Amplification of genes for chorion proteins during oogenesis of Drosophila melanogaster. Proc Natl Acad Sci USA 77:1096–1100

    CAS  PubMed  Google Scholar 

  • Stocker AJ, Pavan C (1974) The influence of ecdysterone on gene amplification, DNA synthesis and puff formation in the salivary gland chromosomes of Rhynchosciara hollanderi. Chromosoma 45:295–319

    CAS  PubMed  Google Scholar 

  • Stocker AJ, Troyano-Pueyo M, Pereira SD, Lara FJS (1984) Ecdysteroid titers and changes in chromosomal activity in the salivary glands of Rhynchosciara americana. Chromosoma 90:26–38

    CAS  Google Scholar 

  • Stocker AJ, Yokosawa J, Soares MA, Cadavid EO (1996) DNA replication and amplification during the final cycle of polyteny in sciarid salivary gland chromosomes and their control by ecdysone. Cienc Cult 48:306–312

    CAS  Google Scholar 

  • Stultz CM, White JV, Smith TF (1993) Structural analysis based on state-space modeling. Protein Sci 2:305–314

    CAS  PubMed  Google Scholar 

  • Stultz CM, Nambudripad R, Lathrop RH, White JV (1997) Predicting protein structure with probabilistic models. In: Allewell N, Woodward C (eds) Protein structural biology in bio-medical research. In: Advances in molecular and cell biology, vol 22B. JAI, Greenwich, pp 447–506

    Google Scholar 

  • Terra WR, de Bianchi AG, Gambarini AG, Pueyo MT, Lara FJS (1976) A medium for short-term incubation of salivary glands based on the haemolymph composition of Rhynchosciara americana. Cienc Cult 28:654–657

    CAS  Google Scholar 

  • Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Winter CE, deBianchi AG, Terra WR, Lara FJS (1977a) Relationships between newly synthesized proteins and DNA puff patterns in salivary glands of Rhynchosciara americana. Chromosoma 61:193–206

    Google Scholar 

  • Winter CE, deBianchi AG, Terra WR, Lara FJS (1977b) The giant DNA puffs of Rhynchosciara americana code for polypeptides of the salivary gland secretion. J Insect Physiol 23:1455–1459

    Article  CAS  Google Scholar 

  • Yokosawa J, Soares MA, Dijkwel PA, Stocker AJ, Hamlin JÁ, Lara FJS (1999) DNA replication of C3 DNA puff amplification initiates at multiples sites in a 6-kb region. Chromosoma 108:291–301

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by FAPESP, CAPES, and CNPq.

Author information

Authors and Affiliations

  1. Depto. Bioquímica, Instituto de Química, Universidade de São Paulo, Cidade Universitária, Av Prof Lineu Prestes, 748, CEP 05508-900, São Paulo, Brazil

    Roberto Vicente Santelli & Fabio Siviero

  2. Depto. Histologia e Embriologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Cidade Universitária, Av Prof Lineu Prestes, 1524, CEP 05508-900, São Paulo, Brazil

    Glaucia Maria Machado-Santelli

  3. Depto. Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, CEP 05508-900, São Paulo, Brazil

    F. J. S. Lara & Ann Jacob Stocker

  4. Centre for Environmental Stress and Adaptation Research, La Trobe University, Bundoora, Victoria, 3086, Australia

    Ann Jacob Stocker

Authors
  1. Roberto Vicente Santelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fabio Siviero
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Glaucia Maria Machado-Santelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. J. S. Lara
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ann Jacob Stocker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Glaucia Maria Machado-Santelli.

Additional information

Communicated by T. Orr-Weaver

This paper is dedicated to the beloved memory of Prof. Dr. Roberto Vicente Santelli.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Santelli, R.V., Siviero, F., Machado-Santelli, G.M. et al. Molecular characterization of the B-2 DNA puff gene of Rhynchosciara americana. Chromosoma 113, 167–176 (2004). https://doi.org/10.1007/s00412-004-0304-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00412-004-0304-6

Keywords

Search

Navigation