DNA methylation and normal chromosome behavior in Neurospora depend on five components of a histone methyltransferase complex, DCDC

PLoS Genet. 2010 Nov 4;6(11):e1001196. doi: 10.1371/journal.pgen.1001196.

Abstract

Methylation of DNA and of Lysine 9 on histone H3 (H3K9) is associated with gene silencing in many animals, plants, and fungi. In Neurospora crassa, methylation of H3K9 by DIM-5 directs cytosine methylation by recruiting a complex containing Heterochromatin Protein-1 (HP1) and the DIM-2 DNA methyltransferase. We report genetic, proteomic, and biochemical investigations into how DIM-5 is controlled. These studies revealed DCDC, a previously unknown protein complex including DIM-5, DIM-7, DIM-9, CUL4, and DDB1. Components of DCDC are required for H3K9me3, proper chromosome segregation, and DNA methylation. DCDC-defective strains, but not HP1-defective strains, are hypersensitive to MMS, revealing an HP1-independent function of H3K9 methylation. In addition to DDB1, DIM-7, and the WD40 domain protein DIM-9, other presumptive DCAFs (DDB1/CUL4 associated factors) co-purified with CUL4, suggesting that CUL4/DDB1 forms multiple complexes with distinct functions. This conclusion was supported by results of drug sensitivity tests. CUL4, DDB1, and DIM-9 are not required for localization of DIM-5 to incipient heterochromatin domains, indicating that recruitment of DIM-5 to chromatin is not sufficient to direct H3K9me3. DIM-7 is required for DIM-5 localization and mediates interaction of DIM-5 with DDB1/CUL4 through DIM-9. These data support a two-step mechanism for H3K9 methylation in Neurospora.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosome Segregation / genetics
  • Chromosomes, Fungal / metabolism*
  • DNA Methylation*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Genes, Essential / genetics
  • Genes, Fungal / genetics
  • Heterochromatin / metabolism
  • Histone Methyltransferases
  • Histone-Lysine N-Methyltransferase / metabolism*
  • Histones / metabolism
  • Lysine / metabolism
  • Models, Biological
  • Multienzyme Complexes / metabolism*
  • Neurospora crassa / enzymology*
  • Neurospora crassa / genetics*
  • Protein Binding

Substances

  • Fungal Proteins
  • Heterochromatin
  • Histones
  • Multienzyme Complexes
  • Histone Methyltransferases
  • Histone-Lysine N-Methyltransferase
  • Lysine