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Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
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AtMCM10 promotes DNA replication-coupled nucleosome assembly in Arabidopsis

Xinjie Zhao1, Jingyi Wang2, Dan Jin1

  • 1State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.

Journal of Integrative Plant Biology
|December 21, 2022
PubMed

Insights

Minichromosome Maintenance protein 10 (MCM10) in Arabidopsis (AtMCM10) binds histones and promotes nucleosome assembly. Loss of AtMCM10 and CAF-1 causes embryo lethality, highlighting their essential roles in DNA replication.

Area of Science:

  • Molecular Biology
  • Epigenetics
  • Plant Science

Background:

  • Minichromosome Maintenance protein 10 (MCM10) is crucial for DNA replication in yeasts and animals.
  • Detailed mechanisms of MCM10 in DNA replication and repair are not fully understood.

Purpose of the Study:

  • Identify and characterize the function of the Arabidopsis MCM10 gene (AtMCM10).
  • Investigate the role of AtMCM10 in DNA replication, nucleosome assembly, and epigenetic regulation.

Main Methods:

  • Forward genetic screening to identify AtMCM10.
  • In vitro binding assays with histone H3-H4.
  • Analysis of nucleosome density in wild-type and Atmcm10 mutant plants.
  • Investigating interactions with Chromatin Assembly Factor-1 (CAF-1).
  • Assessing histone abundance and epigenetic modifications (H3K27me1/3) in Atmcm10 mutants.

Main Results:

  • AtMCM10 is not essential for Arabidopsis growth but binds histone H3-H4 and promotes nucleosome assembly in vitro.
  • Atmcm10 mutants exhibit decreased nucleosome density, partially overlapping with regions regulated by CAF-1.
  • Loss of both AtMCM10 and CAF-1 is embryo lethal, confirming their joint essentiality for replication-coupled nucleosome assembly.
  • Atmcm10 mutants show reduced H3.1 abundance and H3K27me1/3 levels, leading to the release of some silenced transposons.

Conclusions:

  • AtMCM10 plays a vital role in depositing new and parental histones during replication-coupled nucleosome assembly.
  • AtMCM10 is crucial for maintaining epigenetic modifications and genome stability during DNA replication in Arabidopsis.
  • AtMCM10 and CAF-1 function together, highlighting a conserved mechanism for nucleosome assembly during DNA replication.
Keywords:
AtMCM10H3K27me1nucleosomes assembly

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