Control of DEMETER DNA demethylase gene transcription in male and female gamete companion cells in Arabidopsis thaliana

Jin Sup Park, Jennifer M. Frost, Kyunghyuk Park, Hyonhwa Ohr, Guen Tae Park, Seohyun Kim, Hyunjoo Eom, Ilha Lee, Janie S. Brooks, Robert L. Fischer, Yeonhee Choi

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

The DEMETER (DME) DNA glycosylase initiates active DNA demethylation via the base-excision repair pathway and is vital for reproduction in Arabidopsis thaliana. DME-mediated DNA demethylation is preferentially targeted to small, AT-rich, and nucleosome-depleted euchromatic transposable elements, influencing expression of adjacent genes and leading to imprinting in the endosperm. In the female gametophyte, DME expression and subsequent genome-wide DNA demethylation are confined to the companion cell of the egg, the central cell. Here, we show that, in the male gametophyte, DME expression is limited to the companion cell of sperm, the vegetative cell, and to a narrow window of time: immediately after separation of the companion cell lineage from the germline. We define transcriptional regulatory elements of DME using reporter genes, showing that a small region, which surprisingly lies within the DME gene, controls its expression in male and female companion cells. DME expression from this minimal promoter is sufficient to rescue seed abortion and the aberrant DNA methylome associated with the null dme-2 mutation. Within this minimal promoter, we found short, conserved enhancer sequences necessary for the transcriptional activities of DME and combined predicted binding motifs with published transcription factor binding coordinates to produce a list of candidate upstream pathway members in the genetic circuitry controlling DNA demethylation in gamete companion cells. These data show how DNA demethylation is regulated to facilitate endosperm gene imprinting and potential transgenerational epigenetic regulation, without subjecting the germline to potentially deleterious transposable element demethylation.

Original languageEnglish
Pages (from-to)2078-2083
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number8
DOIs
Publication statusPublished - 2017 Feb 21

Bibliographical note

Funding Information:
We thank Jihyun Yu (Seoul National University) for thoughts about the promoter assays and for providing vectors. This study used the Vincent J. Coates Genomics Laboratory at the University of California, Berkeley, supported by NIH S10 Instrumentation Grants S10RR029668 and S10RR027303. This work was supported by Next-Generation BioGreen 21 Program Grant PJ01101802 (to Y.C.), NRF of Korea Grant 2014R1A2A2A01004887 (to Y.C.), and NIH Grant GM69415 (to R.L.F.). J.-S.P. was supported by the Stadelmann-Lee Scholarship Fund, Seoul National University.

Publisher Copyright:
© 2017, National Academy of Sciences. All rights reserved.

All Science Journal Classification (ASJC) codes

  • General

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