---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '19602'
abstract:
- lang: eng
  text: N4-methylcytosine (4mC) is an important DNA modification in prokaryotes, but
    its relevance and even its presence in eukaryotes have been mysterious. Here we
    show that spermatogenesis in the liverwort Marchantia polymorpha involves two
    waves of extensive DNA methylation reprogramming. First, 5-methylcytosine (5mC)
    expands from transposons to the entire genome. Notably, the second wave installs
    4mC throughout genic regions, covering over 50% of CG sites in sperm. 4mC requires
    a methyltransferase (MpDN4MT1a) that is specifically expressed during late spermiogenesis.
    Deletion of MpDN4MT1a alters the sperm transcriptome, causes sperm swimming and
    fertility defects, and impairs post-fertilization development. Our results reveal
    extensive 4mC in a eukaryote, identify a family of eukaryotic methyltransferases,
    and elucidate the biological functions of 4mC in reproductive development, thereby
    expanding the repertoire of functional eukaryotic DNA modifications.
acknowledged_ssus:
- _id: Bio
- _id: ScienComp
acknowledgement: We thank Sir Richard Roberts (NEB) for the kind gift of anti-4mC
  antibodies. We are also grateful to the JIC Small Molecule Mass Spectrometry (Lionel
  Hill) and Chemistry (Martin Rejzek) platforms as well as the High Resolution Metabolomics
  Laboratory (Manfred Beckmann, Aberystwyth University) for their assistance with
  LC-MS. Additionally, we acknowledge the assistance of the JIC Bioimaging Facility
  and ISTA Imaging and Optics Facility for microscopy. Finally, we appreciate the
  High Performance Computing resources provided by the ISTA Scientific Computing Facility
  and Norwich BioScience Institute Partnership Computing Infrastructure. This work
  was funded by a Sainsbury Charitable Foundation studentship (J.W.), a UKRI-BBSRC
  Doctoral Training Partnerships studentship (BBT0087171 to J.T.), a European Research
  Council Starting Grant (“SexMeth” 804981 to J.W., S.X., and X.F.), two Biotechnology
  and Biological Sciences Research Council (BBSRC) grants (BBS0096201 and BBP0135111
  to J.Z., M.V., and X.F.), an EMBO Long Term Fellowship (Y.L.), an ISTA Bridge Fellowship
  (S.X.), and ISTA core funding (Y.Y. and X.F.).
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: James
  full_name: Walker, James
  last_name: Walker
- first_name: Jingyi
  full_name: Zhang, Jingyi
  last_name: Zhang
- first_name: Yalin
  full_name: Liu, Yalin
  last_name: Liu
- first_name: Shujuan
  full_name: Xu, Shujuan
  id: 9724dd9d-f591-11ee-bd51-e97ed0652286
  last_name: Xu
- first_name: Yiming
  full_name: Yu, Yiming
  id: 318e643b-8b61-11ed-b69e-aafa103ec8dd
  last_name: Yu
  orcid: 0000-0002-9919-7282
- first_name: Martin
  full_name: Vickers, Martin
  last_name: Vickers
- first_name: Weizhi
  full_name: Ouyang, Weizhi
  id: fec73395-8b60-11ed-b69e-927fda99c743
  last_name: Ouyang
- first_name: Judit
  full_name: Tálas, Judit
  last_name: Tálas
- first_name: Liam
  full_name: Dolan, Liam
  last_name: Dolan
- first_name: Keiji
  full_name: Nakajima, Keiji
  last_name: Nakajima
- first_name: Xiaoqi
  full_name: Feng, Xiaoqi
  id: e0164712-22ee-11ed-b12a-d80fcdf35958
  last_name: Feng
  orcid: 0000-0002-4008-1234
citation:
  ama: Walker J, Zhang J, Liu Y, et al. Extensive N4 cytosine methylation is essential
    for Marchantia sperm function. <i>Cell</i>. 2025;188(11):2890-2906.e14. doi:<a
    href="https://doi.org/10.1016/j.cell.2025.03.014">10.1016/j.cell.2025.03.014</a>
  apa: Walker, J., Zhang, J., Liu, Y., Xu, S., Yu, Y., Vickers, M., … Feng, X. (2025).
    Extensive N4 cytosine methylation is essential for Marchantia sperm function.
    <i>Cell</i>. Elsevier. <a href="https://doi.org/10.1016/j.cell.2025.03.014">https://doi.org/10.1016/j.cell.2025.03.014</a>
  chicago: Walker, James, Jingyi Zhang, Yalin Liu, Shujuan Xu, Yiming Yu, Martin Vickers,
    Weizhi Ouyang, et al. “Extensive N4 Cytosine Methylation Is Essential for Marchantia
    Sperm Function.” <i>Cell</i>. Elsevier, 2025. <a href="https://doi.org/10.1016/j.cell.2025.03.014">https://doi.org/10.1016/j.cell.2025.03.014</a>.
  ieee: J. Walker <i>et al.</i>, “Extensive N4 cytosine methylation is essential for
    Marchantia sperm function,” <i>Cell</i>, vol. 188, no. 11. Elsevier, p. 2890–2906.e14,
    2025.
  ista: Walker J, Zhang J, Liu Y, Xu S, Yu Y, Vickers M, Ouyang W, Tálas J, Dolan
    L, Nakajima K, Feng X. 2025. Extensive N4 cytosine methylation is essential for
    Marchantia sperm function. Cell. 188(11), 2890–2906.e14.
  mla: Walker, James, et al. “Extensive N4 Cytosine Methylation Is Essential for Marchantia
    Sperm Function.” <i>Cell</i>, vol. 188, no. 11, Elsevier, 2025, p. 2890–2906.e14,
    doi:<a href="https://doi.org/10.1016/j.cell.2025.03.014">10.1016/j.cell.2025.03.014</a>.
  short: J. Walker, J. Zhang, Y. Liu, S. Xu, Y. Yu, M. Vickers, W. Ouyang, J. Tálas,
    L. Dolan, K. Nakajima, X. Feng, Cell 188 (2025) 2890–2906.e14.
corr_author: '1'
date_created: 2025-04-20T22:01:28Z
date_published: 2025-05-29T00:00:00Z
date_updated: 2026-04-28T13:36:51Z
day: '29'
ddc:
- '570'
department:
- _id: XiFe
doi: 10.1016/j.cell.2025.03.014
ec_funded: 1
external_id:
  isi:
  - '001504744800006'
  pmid:
  - '40209706'
file:
- access_level: open_access
  checksum: 0dcc2feb368dfe7c4890093366b2dacb
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  creator: dernst
  date_created: 2025-12-29T13:40:32Z
  date_updated: 2025-12-29T13:40:32Z
  file_id: '20871'
  file_name: 2025_Cell_Walker.pdf
  file_size: 11622960
  relation: main_file
  success: 1
file_date_updated: 2025-12-29T13:40:32Z
has_accepted_license: '1'
intvolume: '       188'
isi: 1
issue: '11'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '05'
oa: 1
oa_version: Published Version
page: 2890-2906.e14
pmid: 1
project:
- _id: bdb51a6e-d553-11ed-ba76-c2025f3d5725
  call_identifier: H2020
  grant_number: '804981'
  name: Establishment, modulation and inheritance of sexual lineage specific DNA methylation
    in plants
publication: Cell
publication_identifier:
  eissn:
  - 1097-4172
  issn:
  - 0092-8674
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  link:
  - description: News on ISTA website
    relation: press_release
    url: https://ista.ac.at/en/news/from-bacterial-immunity-to-plant-sex/
scopus_import: '1'
status: public
title: Extensive N4 cytosine methylation is essential for Marchantia sperm function
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
volume: 188
year: '2025'
...
---
OA_place: publisher
OA_type: hybrid
_id: '15375'
abstract:
- lang: eng
  text: In the eukaryotic nucleus, heterochromatin forms highly condensed, visible
    foci known as heterochromatin foci (HF). These HF are enriched with linker histone
    H1, a key player in heterochromatin condensation and silencing. However, it is
    unknown how H1 aggregates HF and condenses heterochromatin. In this study, we
    established that H1 facilitates heterochromatin condensation by enhancing inter-
    and intrachromosomal interactions between and within heterochromatic regions of
    the Arabidopsis (Arabidopsis thaliana) genome. We demonstrated that H1 drives
    HF formation via phase separation, which requires its C-terminal intrinsically
    disordered region (C-IDR). A truncated H1 lacking the C-IDR fails to form foci
    or recover HF in the h1 mutant background, whereas C-IDR with a short stretch
    of the globular domain (18 out of 71 amino acids) is sufficient to rescue both
    defects. In addition, C-IDR is essential for H1's roles in regulating nucleosome
    repeat length and DNA methylation in Arabidopsis, indicating that phase separation
    capability is required for chromatin functions of H1. Our data suggest that bacterial
    H1-like proteins, which have been shown to condense DNA, are intrinsically disordered
    and capable of mediating phase separation. Therefore, we propose that phase separation
    mediated by H1 or H1-like proteins may represent an ancient mechanism for condensing
    chromatin and DNA.
acknowledged_ssus:
- _id: Bio
- _id: ScienComp
acknowledgement: "This work was funded by ISTA core support (Y.Y. and X.F.) and grants
  from the National Natural Science Foundation of China (31871443 to L.W. and P.L.;
  32100417 to L.W.).\r\nWe thank the ISTA Imaging and Optics Facility for assistance
  with microscopy and the ISTA Scientific Computing Facility for high-performance
  computing resources."
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Shengbo
  full_name: He, Shengbo
  last_name: He
- first_name: Yiming
  full_name: Yu, Yiming
  id: 318e643b-8b61-11ed-b69e-aafa103ec8dd
  last_name: Yu
- first_name: Liang
  full_name: Wang, Liang
  last_name: Wang
- first_name: Jingyi
  full_name: Zhang, Jingyi
  last_name: Zhang
- first_name: Zhengyong
  full_name: Bai, Zhengyong
  last_name: Bai
- first_name: Guohong
  full_name: Li, Guohong
  last_name: Li
- first_name: Pilong
  full_name: Li, Pilong
  last_name: Li
- first_name: Xiaoqi
  full_name: Feng, Xiaoqi
  id: e0164712-22ee-11ed-b12a-d80fcdf35958
  last_name: Feng
  orcid: 0000-0002-4008-1234
citation:
  ama: He S, Yu Y, Wang L, et al. Linker histone H1 drives heterochromatin condensation
    via phase separation in Arabidopsis. <i>The Plant Cell</i>. 2024;36(5):1829-1843.
    doi:<a href="https://doi.org/10.1093/plcell/koae034">10.1093/plcell/koae034</a>
  apa: He, S., Yu, Y., Wang, L., Zhang, J., Bai, Z., Li, G., … Feng, X. (2024). Linker
    histone H1 drives heterochromatin condensation via phase separation in Arabidopsis.
    <i>The Plant Cell</i>. Oxford University Press. <a href="https://doi.org/10.1093/plcell/koae034">https://doi.org/10.1093/plcell/koae034</a>
  chicago: He, Shengbo, Yiming Yu, Liang Wang, Jingyi Zhang, Zhengyong Bai, Guohong
    Li, Pilong Li, and Xiaoqi Feng. “Linker Histone H1 Drives Heterochromatin Condensation
    via Phase Separation in Arabidopsis.” <i>The Plant Cell</i>. Oxford University
    Press, 2024. <a href="https://doi.org/10.1093/plcell/koae034">https://doi.org/10.1093/plcell/koae034</a>.
  ieee: S. He <i>et al.</i>, “Linker histone H1 drives heterochromatin condensation
    via phase separation in Arabidopsis,” <i>The Plant Cell</i>, vol. 36, no. 5. Oxford
    University Press, pp. 1829–1843, 2024.
  ista: He S, Yu Y, Wang L, Zhang J, Bai Z, Li G, Li P, Feng X. 2024. Linker histone
    H1 drives heterochromatin condensation via phase separation in Arabidopsis. The
    Plant Cell. 36(5), 1829–1843.
  mla: He, Shengbo, et al. “Linker Histone H1 Drives Heterochromatin Condensation
    via Phase Separation in Arabidopsis.” <i>The Plant Cell</i>, vol. 36, no. 5, Oxford
    University Press, 2024, pp. 1829–43, doi:<a href="https://doi.org/10.1093/plcell/koae034">10.1093/plcell/koae034</a>.
  short: S. He, Y. Yu, L. Wang, J. Zhang, Z. Bai, G. Li, P. Li, X. Feng, The Plant
    Cell 36 (2024) 1829–1843.
corr_author: '1'
date_created: 2024-05-12T22:01:01Z
date_published: 2024-05-01T00:00:00Z
date_updated: 2025-09-08T07:21:17Z
day: '01'
ddc:
- '580'
department:
- _id: XiFe
doi: 10.1093/plcell/koae034
external_id:
  isi:
  - '001180817000001'
  pmid:
  - '38309957'
file:
- access_level: open_access
  checksum: eed76c848fe3d8fe9a53943181aaa53c
  content_type: application/pdf
  creator: dernst
  date_created: 2025-04-23T07:43:12Z
  date_updated: 2025-04-23T07:43:12Z
  file_id: '19611'
  file_name: 2024_PlantCell_He.pdf
  file_size: 50791962
  relation: main_file
  success: 1
file_date_updated: 2025-04-23T07:43:12Z
has_accepted_license: '1'
intvolume: '        36'
isi: 1
issue: '5'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
page: 1829-1843
pmid: 1
publication: The Plant Cell
publication_identifier:
  eissn:
  - 1532-298X
publication_status: published
publisher: Oxford University Press
quality_controlled: '1'
scopus_import: '1'
status: public
title: Linker histone H1 drives heterochromatin condensation via phase separation
  in Arabidopsis
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 36
year: '2024'
...