Internuclear gene silencing in Phytophthora infestans is established through chromatin remodelling

Pieter van West, Samantha J. Shepherd, Claire A. Walker, Shuang Li, Alex A. Appiah, Laura J. Grenville-Briggs, Francine Govers, Neil A. R. Gow

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

In the plant pathogen Phytophthora intestans, nuclear integration of inf1 transgenic DNA sequences results in internuclear gene silencing of inf1. Although silencing is regulated at the transcriptional level, it also affects transcription from other nuclei within heterokaryotic cells of the mycelium. Here we report experiments exploring the mechanism of internuclear gene silencing in P. infestans. The DNA methylation inhibitor 5-azacytidine induced reversion of the inf1-silenced state. Also, the histone deacetylase inhibitor trichostatin-A was able to reverse inf1 silencing. inf1-expression levels returned to the silenced state when the inhibitors were removed except in non-transgenic inf1-silenced strains that were generated via internuclear gene silencing, where inf1 expression was restored permanently. Therefore, inf1-transgenic sequences are required to maintain the silenced state. Prolonged culture of non-transgenic inf1-silenced strains resulted in gradual reactivation of inf1 gene expression. Nuclease digestion of inf1-silenced and non-silenced nuclei showed that inf1 sequences in silenced nuclei were less rapidly degraded than non-silenced inf1 sequences. Bisulfite sequencing of the endogenous inf1 locus did not result in detection of any cytosine methylation. Our findings suggest that the inf1-silenced state is based on chromatin remodelling.

Original languageEnglish
Pages (from-to)1482-1490
Number of pages9
JournalMicrobiology
Volume154
Issue number5
DOIs
Publication statusPublished - May 2008

Keywords

  • double-stranded-RNA
  • global histone acetylation
  • DNA methylation
  • transcriptional repression
  • caenorhabditis-elegans
  • filamentous fungi
  • transgene
  • plants
  • interference
  • maintenenance

Cite this

van West, P., Shepherd, S. J., Walker, C. A., Li, S., Appiah, A. A., Grenville-Briggs, L. J., Govers, F., & Gow, N. A. R. (2008). Internuclear gene silencing in Phytophthora infestans is established through chromatin remodelling. Microbiology , 154(5), 1482-1490. https://doi.org/10.1099/mic.0.2007/015545-0