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Current Genetics

, Volume 43, Issue 6, pp 425–432 | Cite as

Unpaired genes do not silence their paired neighbors

  • Brandi L. Kutil
  • Kye-Yong Seong
  • Rodolfo AramayoEmail author
Research Article

Abstract

During meiotic chromosome pairing, a loop of unpaired DNA induces the silencing of all paired and unpaired homologous DNA via meiotic silencing, an RNA-mediated post-transcriptional gene-silencing mechanism. To test the effect of unpaired DNA on adjacent genes, we constructed strains containing the DNA of a transformation marker integrated immediately downstream of the Ascospore maturation-1 (Asm-1) gene and tested whether this unpaired DNA silences asm-1+. We conclude that unpaired downstream DNA has no effect on Asm-1 expression during meiosis or ascospore development, which suggests that the silencing signal produced by unpaired DNA does not propagate onto adjacent paired regions.

Keywords

Chromosome pairing RNA silencing Meiotic silencing Gene replacement Locus engineering 

Notes

Acknowledgements

We thank Robert J. Pratt for his critical review of the manuscript. This work was supported by United States Public Health Service grant GM58770 to R.A.

References

  1. Aramayo R, Metzenberg RL (1996) Meiotic transvection in fungi. Cell 86:103–113PubMedGoogle Scholar
  2. Aramayo R, Peleg Y, Addison R, Metzenberg R (1996) Asm-1 +, a Neurospora crassa gene related to transcriptional regulators of fungal development. Genetics 144:991–1003PubMedGoogle Scholar
  3. Ausubel FM, et al (1987) Current protocols in molecular biology. Wiley, New YorkGoogle Scholar
  4. Carroll AM, Sweigard JA, Valent V (1994) Improved vectors for selecting resistance to hygromycin. Fungal Genet Newsl 41:22Google Scholar
  5. Davis RH, Serres FJ de (1970) Genetic and microbiological research techniques for Neurospora crassa. In: Colowick SP, Kaplan NO (eds) Metabolism of amino acids and amines. Academic Press, New York, pp 79–143Google Scholar
  6. Lee DW, Haag JR, Aramayo R (2003a) Construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 (his-3) locus of Neurospora crassa. Curr Genet 43:17–23Google Scholar
  7. Lee DW, Pratt RJ, McLaughin M, Aramayo R (2003b) An Argonaute-like protein is required for meiotic silencing. Genetics 164 (2)Google Scholar
  8. Lupton SD, Brunton LL, Kalberg VA, Overell RW (1991) Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene. Mol Cell Biol 11:3374–3378PubMedGoogle Scholar
  9. Marsh JL, Erfle M, Wykes EJ (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32:481–485PubMedGoogle Scholar
  10. Pratt RJ, Aramayo R (2002) Improving the efficiency of gene replacements in Neurospora crassa: a first step towards a large-scale functional genomics project. Fungal Genet Biol 37:56–71CrossRefPubMedGoogle Scholar
  11. Raju NB (1980) Meiosis and ascospore genesis in Neurospora. Eur J Cell Biol 23:208–223PubMedGoogle Scholar
  12. Raju NB (1992) Genetic control of the sexual cycle in Neurospora. Mycol Res 96:241–262Google Scholar
  13. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  14. Shiu PK, Metzenberg RL (2002) Meiotic silencing by unpaired DNA. Properties, regulation and suppression. Genetics 161:1483–1495PubMedGoogle Scholar
  15. Shiu PKT, Raju BN, Zickler D, Metzenberg R (2001) Meiotic silencing by unpaired DNA. Cell 107:905–916PubMedGoogle Scholar
  16. Springer ML (1993) Genetic control of fungal differentiation: the three sporulation pathways of Neurospora crassa. BioEssays 15:365–374PubMedGoogle Scholar
  17. Staben C, et al (1989) Use of a bacterial hygromycin B resistance gene as a dominant selectable marker in Neurospora crassa transformation. Fungal Genet Newsl 36:79–81Google Scholar
  18. Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119PubMedGoogle Scholar
  19. Zickler D, Kleckner N (1998) The leptotene–zygotene transition of meiosis. Annu Rev Genet 32:619–697PubMedGoogle Scholar
  20. Zickler D, Kleckner N (1999) Meiotic chromosomes: integrating structure and function. Annu Rev Genet 33:603–754PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Brandi L. Kutil
    • 2
  • Kye-Yong Seong
    • 1
  • Rodolfo Aramayo
    • 1
    Email author
  1. 1.Department of Biology, College of ScienceTexas A&M UniversityCollege StationUSA
  2. 2.Department of Plant Pathology and Microbiology, College of AgricultureTexas A&M UniversityCollege StationUSA

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