The multi-domain protein Np95 connects DNA methylation and histone modification.
DNA methylation and histone modifications play a central role in the epigenetic regulation of gene expression and cell differentiation. Recently, Np95 (also known as UHRF1 or ICBP90) has been found to interact with Dnmt1 and to bind hemimethylated DNA, indicating together with genetic studies a central role in the maintenance of DNA methylation. Using in vitro binding assays we observed a weak preference of Np95 and its SRA (SET- and Ring-associated) domain for hemimethylated CpG sites. However, the binding kinetics of Np95 in living cells was not affected by the complete loss of genomic methylation. Investigating further links with heterochromatin, we could show that Np95 preferentially binds histone H3 N-terminal tails with trimethylated (H3K9me3) but not acetylated lysine 9 via a tandem Tudor domain. This domain contains three highly conserved aromatic amino acids that form an aromatic cage similar to the one binding H3K9me3 in the chromodomain of HP1ss. Mutations targeting the aromatic cage of the Np95 tandem Tudor domain (Y188A and Y191A) abolished specific H3 histone tail binding. These multiple interactions of the multi-domain protein Np95 with hemimethylated DNA and repressive histone marks as well as with DNA and histone methyltransferases integrate the two major epigenetic silencing pathways.
Rottach
Andrea
A
Ludwig Maximilians University Munich, Department of Biology II and Center for Integrated Protein Science Munich, Grosshaderner Str. 2, 82152 Planegg-Martinsried, Germany.
Frauer
Carina
C
Pichler
Garwin
G
Bonapace
Ian Marc
IM
Spada
Fabio
F
Leonhardt
Heinrich
H
D000595:Amino Acid Sequence; D022762:CCAAT-Enhancer-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D019175:DNA Methylation; D006657:Histones / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D044767:Ubiquitin-Protein Ligases
D022762:CCAAT-Enhancer-Binding Proteins; D006657:Histones; D004247:DNA; C406364:UHRF1 protein, human; D044767:Ubiquitin-Protein Ligases