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Methylation of the foreign transposon Restless in vegetative mycelia of Neurospora crassa

Abstract

Methylation of foreign and/or repeated sequences in the filamentous fungus Neurospora crassa is believed to be directed against invading transposable elements. To test this hypothesis, the fate of a transposon in N. crassa was investigated. Vectors were constructed which carried the transposon Restless, an active class-II element isolated from the fungus Tolypocladium inflatum. These vectors were introduced into N. crassa strains by protoplast transformation. Two strategies were employed: (1) ectopic multi-copy integration, and (2) site-specific single-copy integration at the his-3 locus. All ectopic transformants exhibited strong methylation as confirmed by Southern hybridization of genomic DNA digested with the methylation-sensitive endonuclease Sau3AI and the methylation-insensitive endonuclease NdeII. Single copies of Restless integrated at the his-3 locus were not methylated. These results are discussed with respect to non-RIP methylation and potential consequences for gene-tagging strategies based on the use of Restless.

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Similar content being viewed by others Explore related subjectsDiscover the latest articles and news from researchers in related subjects, suggested using machine learning. Author information Authors and Affiliations
  1. Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, D-44780 Bochum, Germany e-mail: frank.kempken@ruhr-uni-bochum.de Tel.: +49 (0)234/32-22465 Fax: +49 (0)234-32-14184, , , , , , DE

    Frank Windhofer & Frank Kempken

  2. Flinders University, School of Biological Sciences, PO Box 2100, Adelaide SA 5001, Australia, , , , , , AU

    David E. A. Catcheside

Authors
  1. Frank Windhofer
  2. David E. A. Catcheside
  3. Frank Kempken
Additional information

Received: 27 September / 3 November 1999

About this article Cite this article

Windhofer, F., Catcheside, D. & Kempken, F. Methylation of the foreign transposon Restless in vegetative mycelia of Neurospora crassa . Curr Genet 37, 194–199 (2000). https://doi.org/10.1007/s002940050519

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