PpCMT chromomethylase affects cell growth and interacts with the homolog of LIKE HETEROCHROMATIN PROTEIN 1 in the moss Physcomitrella patens

• Published in: The Plant journal : for cell and molecular biology 2014-02, Vol.77 (4), p.589-603
• Main Authors: Dangwal, Meenakshi, Kapoor, Sanjay, Kapoor, Meenu
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bryopsida - cytology; Bryopsida - enzymology; Bryopsida - genetics; Bryopsida - growth & development; cell growth; cell walls; chromomethylase; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; cytosine; Cytosine - metabolism; Dimerization; DNA; DNA (Cytosine-5-)-Methyltransferases - genetics; DNA (Cytosine-5-)-Methyltransferases - metabolism; DNA Methylation; DNA, Plant - genetics; Enzymes; fluorescence; Gene expression; Gene Expression Regulation, Plant; Gene Silencing; genes; Genome, Plant - genetics; Germ Cells, Plant; heterochromatin; histones; LIKE HETEROCHROMATIN PROTEIN 1; metabolism; methyltransferases; Models, Molecular; Molecular Sequence Data; mosses and liverworts; mutants; Mutation; Paleobotany; Phylogeny; Physcomitrella patens; Plant biology; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified; polymerase chain reaction; Protein Structure, Tertiary; Proteins; protonemata; Sequence Alignment; Spermatophyta; Two-Hybrid System Techniques; xyloglucans; yeast two‐hybrid; yeasts
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/tpj.12406

Chromomethylases (CMTs) are plant‐specific cytosine DNA methyltransferases that are involved in maintenance of CpNpG methylation. In seed plants, histone methylation and interaction of CMT with LIKE HETEROCHROMATIN PROTEIN 1 (LHP1) is essential for recruitment of CMT to target sites. LHP1 has been characterized as a putative component of the POLYCOMB REPRESSIVE COMPLEX1 (PRC1) in plants, and functions downstream of PRC2 to maintain genes in repressed state for orchestrated development. In the present study, we show that targeted disruption of PpCMT results in an approximately 50% reduction in global cytosine methylation levels. This affects growth of apical cells, predominantly growth of side branch initials emerging from chloronema cells. In some places, these cells develop thick walls with plasmolyzed cellular contents. Transcript accumulation patterns of genes involved in apical cell extension and metabolism of hemicelluloses, such as xyloglucans, in the primary cell walls decreased many fold in ppcmt mutant lines, as determined by real‐time PCR. Using yeast two‐hybrid method and bimolecular fluorescence complementation assay, we show that PpCMT and PpLHP1 interact through their chromo domains, while PpLHP1 homodimerizes through its chromo shadow domain. The results presented in this study provide insight into the role of the single chromomethylase, PpCMT, in proliferation of protonema filaments, and shed light on the evolutionary conservation of proteins interacting with these methylases in the early land plant, Physcomitrella patens.