DCAF26, an adaptor protein of Cul4-based E3, is essential for DNA methylation in Neurospora crassa.
DNA methylation is involved in gene silencing and genome stability in organisms from fungi to mammals. Genetic studies in Neurospora crassa previously showed that the CUL4-DDB1 E3 ubiquitin ligase regulates DNA methylation via histone H3K9 trimethylation. However, the substrate-specific adaptors of this ligase that are involved in the process were not known. Here, we show that, among the 16 DDB1- and Cul4-associated factors (DCAFs) encoded in the N. crassa genome, three interacted strongly with CUL4-DDB1 complexes. DNA methylation analyses of dcaf knockout mutants revealed that dcaf26 was required for all of the DNA methylation that we observed. In addition, histone H3K9 trimethylation was also eliminated in dcaf26(KO) mutants. Based on the finding that DCAF26 associates with DDB1 and the histone methyltransferase DIM-5, we propose that DCAF26 protein is the major adaptor subunit of the Cul4-DDB1-DCAF26 complex, which recruits DIM-5 to DNA regions to initiate H3K9 trimethylation and DNA methylation in N. crassa.
Xu
Hui
H
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
Wang
Jiyong
J
Hu
Qiwen
Q
Quan
Yun
Y
Chen
Huijie
H
Cao
Yingqiong
Y
Li
Chunbo
C
Wang
Ying
Y
He
Qun
Q
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D044842:Cullin Proteins / Q000378:metabolism*; D019175:DNA Methylation; D004271:DNA, Fungal / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D055786:Gene Knockout Techniques; D005800:Genes, Fungal / Q000235:genetics; D006657:Histones / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D016415:Sequence Alignment; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D044842:Cullin Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins; D006657:Histones; D044767:Ubiquitin-Protein Ligases; D008239:Lysine