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A single CMT methyltransferase homolog is involved in CHG DNA methylation and development of Physcomitrella patens

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Abstract

C-5 DNA methylation is an essential mechanism controlling gene expression and developmental programs in a variety of organisms. Though the role of DNA methylation has been intensively studied in mammals and Arabidopsis, little is known about the evolution of this mechanism. The chromomethylase (CMT) methyltransferase family is unique to plants and was found to be involved in DNA methylation in Arabidopsis, maize and tobacco. The moss Physcomitrella patens, a model for early terrestrial plants, harbors a single homolog of the CMT protein family designated as PpCMT. Our phylogenetic analysis suggested that the CMT family is unique to embryophytes and its earliest known member PpCMT belongs to the CMT3 subfamily. Thus, P. patens may serve as a model to study the ancient functions of the CMT3 family. We have generated a ΔPpcmt deletion mutant which demonstrated that PpCMT is essential for P. patens protonema and gametophore development and is involved in CHG methylation as demonstrated at four distinct genomic loci. PpCMT protein accumulation pattern correlated with proliferating cells and was sub-localized to the nucleus as predicted from its function. Taken together, our results suggested that CHG DNA methylation mediated by CMT has been employed early in land plant evolution to control developmental programs during both the vegetative and reproductive haploid phases along the plant life cycle.

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Abbreviations

BAH:

Bromo-adjacent homology domain

CMT:

Chromomethylase

DNMT:

DNA methyltransferase

DRM:

Domains rearranged methyltransferase

GUS:

β-Glucuronidase

MTD:

C-5 DNA methyltransferase domain

MET1:

DNA methyltransferase 1

RdDM:

RNA directed DNA methylation

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Acknowledgments

We thank Professors Tal Pupko, Eran Bacharach, Shaul Yalovsky and Daniel Chamovitz for critical reading and helpful comments. C.N.M and R.Y were supported in part by a matching Tel-Aviv University Deans doctoral fellowship and the Manna foundation. This research was supported by the Israeli Science Foundation Grant #767/09, and by the Israel Korea program # 3-824 financed by the Ministry of Science and Technology, both granted to N.O. Transgenic lines described in this study were deposited in the International Moss Stock Center (http://www.moss-stock-center.org/) with the accessions IMSC 40736, 40737, 40738 (ΔPpcmt 103, 157 and 281) and 40739, 40740 (PpCMT–GUS 7 and 8).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Molecular Biology and Ecology of Plants, Tel-Aviv University, 69978, Tel-Aviv, Israel

    Chen Noy-Malka, Rafael Yaari, Rachel Itzhaki, Nitzan Auerbach Gershovitz, Aviva Katz & Nir Ohad

  2. Department of Plant Sciences, The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Hebrew University of Jerusalem, 76100, Rehovot, Israel

    Assaf Mosquna

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  1. Chen Noy-Malka
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  2. Rafael Yaari
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  3. Rachel Itzhaki
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  4. Assaf Mosquna
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  6. Aviva Katz
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Correspondence to Aviva Katz or Nir Ohad.

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Chen Noy-Malka and Rafael Yaari have contributed equally to the work.

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Noy-Malka, C., Yaari, R., Itzhaki, R. et al. A single CMT methyltransferase homolog is involved in CHG DNA methylation and development of Physcomitrella patens . Plant Mol Biol 84, 719–735 (2014). https://doi.org/10.1007/s11103-013-0165-6

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