The RFTS domain of Raf2 is required for Cul4 interaction and heterochromatin integrity in fission yeast

Autor: White, SA, Buscaino, A, Sanchez-Pulido, L, Ponting, C, Nowicki, M, Allshire, R
Jazyk: angličtina
Rok vydání: 2014
Předmět:
DNA (Cytosine-5-)-Methyltransferase 1
Chromosome Structure and Function
Science
Centromere
Mitosis
Chromosomes
Molecular Genetics
Schizosaccharomyces Pombe
RNA interference
Chromosome Segregation
Heterochromatin
Schizosaccharomyces
Genetics
Humans
Point Mutation
Fungal Genetics
DNA (Cytosine-5-)-Methyltransferases
RNA
Small Interfering
Molecular Biology
Centromeres
Medicine(all)
Biology and life sciences
Agricultural and Biological Sciences(all)
Chromosome Biology
Biochemistry
Genetics and Molecular Biology(all)
Organisms
Fungi
Cell Biology
Histone-Lysine N-Methyltransferase
DNA Methylation
Cullin Proteins
Chromatin
Yeast
Protein Structure
Tertiary
DNA (Cytosine-5-)-Methyltransferase
Histone Methyltransferases
Medicine
Epigenetics
Schizosaccharomyces pombe Proteins
Histone modification
Sequence Alignment
Research Article
Zdroj: White, S A, Buscaino, A, Sanchez-Pulido, L, Ponting, C P, Nowicki, M W & Allshire, R C 2014, ' The RFTS domain of Raf2 is required for Cul4 interaction and heterochromatin integrity in fission yeast ', PLoS ONE, vol. 9, no. 8, e104161, pp. e104161 . https://doi.org/10.1371/journal.pone.0104161
PLoS ONE, Vol 9, Iss 8, p e104161 (2014)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0104161
Popis: Centromeric heterochromatin assembly in fission yeast is critical for faithful chromosome segregation at mitosis. Its assembly requires a concerted pathway of events whereby the RNA interference (RNAi) pathway guides H3K9 methylation to target sequences. H3K9 methylation, a hallmark of heterochromatin structure, is mediated by the single histone methyltransferase Clr4 (equivalent to metazoan Suv3-9), a component of the CLRC complex. Loss of or defects in CLRC components disrupts heterochromatin formation due to loss of H3K9 methylation, thus an intact, fully functional CLRC complex is required for heterochromatin integrity. Despite its importance, little is known about the contribution of the CLRC component Raf2 to H3K9 methylation and heterochromatin assembly. We demonstrate that Raf2 is concentrated at centromeres and contrary to other analyses, we find that loss of Raf2 does not affect CENP-A Cnp1 localisation or recruitment to centromeres. Our sequence alignments show that Raf2 contains a Replication Foci Targeting Sequence (RFTS) domain homologous to the RFTS domain of the human DNA methyltransferase DNMT1. We show that the Raf2 RFTS domain is required for centromeric heterochromatin formation as its mutation disrupts H3K9 methylation but not the processing of centromeric transcripts into small interfering RNAs (siRNAs) by the RNAi pathway. Analysis of biochemical interactions demonstrates that the RFTS domain mediates an interaction between Raf2 and the CLRC component Cul4. We conclude that the RFTS domain of Raf2 is a protein interaction module that plays an important role in heterochromatin formation at centromeres.
Databáze: OpenAIRE
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