Feb 25, 2016

Histone H1 limits DNA methylation in Neurospora crassa

BioRxiv : the Preprint Server for Biology
Michael SeymourZachary A Lewis

Abstract

Histone H1 variants, known as linker histones, are essential chromatin components in higher eukaryotes, yet compared to the core histones relatively little is known about their in vivo functions. The filamentous fungus Neurospora crassa encodes a single H1 protein that is not essential for viability. To investigate the role of N. crassa H1, we constructed a functional FLAG-tagged H1 fusion protein and performed genomic and molecular analyses. Cell fractionation experiments showed that H1-FLAG is a chromatin binding protein. Chromatin-immunoprecipitation combined with sequencing (ChIP-seq) revealed that H1-3XFLAG is globally enriched throughout the genome with a subtle preference for promoters of expressed genes. In mammals, the stochiometery of H1 impacts nucleosome repeat length. To determine if H1 impacts nucleosome occupancy or nucleosome positioning in N. crassa, we performed Micrococcal nuclease digestion in wildtype and the ∆hH1 strain followed by sequencing (MNase-seq). Deletion of hH1 did not significantly impact nucleosome positioning or nucleosome occupancy. Analysis of DNA methylation by whole-genome bisulfite sequencing (MethylC-seq) revealed a modest but global increase in DNA methylation in the ∆hH1 mutant. Togeth...Continue Reading

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Mentioned in this Paper

SCN5A protein, human
Neurospora crassa
In Vivo
Hydrogen sulfite
DNA Methylation [PE]
Nucleosomes
Micrococcal Nuclease
Genome
Genes
Filamentous fungus

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