Seymour, Michael, Ji, Lexiang, Santos, Alex M, Kamei, Masayuki, Sasaki, Takahiko, Basenko, Evelina Y 
ORCID: 0000-0001-8611-5447, Schmitz, Robert J, Zhang, Xiaoyu and Lewis, Zachary A
(2016)
Histone H1 Limits DNA Methylation in <i>Neurospora crassa</i>.
G3-GENES GENOMES GENETICS, 6 (7).
pp. 1879-1889.
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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 H1-3XFLAG 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 stoichiometry 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 the wild-type and the [Formula: see text]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 [Formula: see text]hH1 mutant. Together, these data suggest that H1 acts as a nonspecific chromatin binding protein that can limit accessibility of the DNA methylation machinery in N. crassa.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | chromatin, histone H1, nucleosome positioning, DNA methylation |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 28 Sep 2021 09:36 |
| Last Modified: | 16 Oct 2023 17:53 |
| DOI: | 10.1534/g3.116.028324 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3138567 |
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