The MBD7 complex promotes expression of methylated transgenes without significantly altering their methylation status

  1. Dongming Li
  2. Ana Marie S Palanca
  3. So Youn Won
  4. Lei Gao
  5. Ying Feng
  6. Ajay A Vashisht
  7. Li Liu
  8. Yuanyuan Zhao
  9. Xigang Liu
  10. Xiuyun Wu
  11. Shaofang Li
  12. Brandon Le
  13. Yun Ju Kim
  14. Guodong Yang
  15. Shengben Li
  16. Jinyuan Liu
  17. James A Wohlschlegel
  18. Hongwei Guo
  19. Beixin Mo
  20. Xuemei Chen  Is a corresponding author
  21. Julie A Law  Is a corresponding author
  1. University of California, Riverside, United States
  2. Salk Institute for Biological Studies, United States
  3. Rural Development Administration, Republic of Korea
  4. David Geffen School of Medicine at UCLA, United States
  5. Tsinghua University, China
  6. Peking University, China
  7. Shenzhen University, China

Abstract

DNA methylation is associated with gene silencing in eukaryotic organisms. Although pathways controlling the establishment, maintenance and removal of DNA methylation are known, relatively little is understood about how DNA methylation influences gene expression. Here we identified a METHYL-CpG-BINDING DOMAIN 7 (MBD7) complex in Arabidopsis thaliana that suppresses the transcriptional silencing of two LUCIFERASE (LUC) reporters via a mechanism that is largely downstream of DNA methylation. Although mutations in components of the MBD7 complex resulted in modest increases in DNA methylation concomitant with decreased LUC expression, we found that these hyper-methylation and gene expression phenotypes can be genetically uncoupled. This finding, along with genome-wide profiling experiments showing minimal changes in DNA methylation upon disruption of the MBD7 complex, places the MBD7 complex amongst a small number of factors acting downstream of DNA methylation. This complex, however, is unique as it functions to suppress, rather than enforce, DNA methylation-mediated gene silencing.

Data availability

The following data sets were generated

Article and author information

Author details

  1. Dongming Li

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Ana Marie S Palanca

    Plant Molecular and Cellular Biology Laboratory, Salk Institute for Biological Studies, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. So Youn Won

    National Academy of Agricultural Science, Rural Development Administration, Suwon, Republic of Korea
    Competing interests
    The authors declare that no competing interests exist.
  4. Lei Gao

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Ying Feng

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Ajay A Vashisht

    Department of Biological Chemistry, David Geffen School of Medicine at UCLA, Los Angeles, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Li Liu

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Yuanyuan Zhao

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Xigang Liu

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Xiuyun Wu

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Shaofang Li

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Brandon Le

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Yun Ju Kim

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Guodong Yang

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Shengben Li

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Jinyuan Liu

    Laboratory of Molecular Biology and Protein Science, Tsinghua University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  17. James A Wohlschlegel

    Department of Biological Chemistry, David Geffen School of Medicine at UCLA, Los Angeles, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. Hongwei Guo

    State Key Laboratory of Protein and Plant Gene research, Peking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  19. Beixin Mo

    College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, China
    Competing interests
    The authors declare that no competing interests exist.
  20. Xuemei Chen

    Department of Botany and Plant Sciences, University of California, Riverside, Riverside, United States
    For correspondence
    xuemei.chen@ucr.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5209-1157
  21. Julie A Law

    Plant Molecular and Cellular Biology Laboratory, Salk Institute for Biological Studies, La Jolla, United States
    For correspondence
    jlaw@salk.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7472-7753

Funding

Gordon and Betty Moore Foundation (GBMF3046)

  • Xuemei Chen

National Science Foundation of China (91440105)

  • Xuemei Chen

National Science Foundation of China (30970265)

  • Beixin Mo

National Science Foundation of China (31210103901)

  • Beixin Mo

Guangdong Innovation Research Team Fund (2014ZT05S078)

  • Xuemei Chen

National Institutes of Health (GM061146)

  • Xuemei Chen

National Academy of Agricultural Science (PJ008725)

  • So Youn Won

China Scholarship Council

  • Dongming Li

Glenn Center for Aging Research at the Salk Institute

  • Ana Marie S Palanca

Helmsley Charitable Trust

  • Ana Marie S Palanca
  • Julie A Law

National Institutes of Health (GM112966)

  • Julie A Law

National Institutes of Health (GM089778)

  • James A Wohlschlegel

National Institutes of Health (P30 014195)

  • Ana Marie S Palanca
  • Julie A Law

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2017, Li et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,859
    views
  • 755
    downloads
  • 19
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Dongming Li
  2. Ana Marie S Palanca
  3. So Youn Won
  4. Lei Gao
  5. Ying Feng
  6. Ajay A Vashisht
  7. Li Liu
  8. Yuanyuan Zhao
  9. Xigang Liu
  10. Xiuyun Wu
  11. Shaofang Li
  12. Brandon Le
  13. Yun Ju Kim
  14. Guodong Yang
  15. Shengben Li
  16. Jinyuan Liu
  17. James A Wohlschlegel
  18. Hongwei Guo
  19. Beixin Mo
  20. Xuemei Chen
  21. Julie A Law
(2017)
The MBD7 complex promotes expression of methylated transgenes without significantly altering their methylation status
eLife 6:e19893.
https://doi.org/10.7554/eLife.19893

Share this article

https://doi.org/10.7554/eLife.19893

Further reading

    1. Chromosomes and Gene Expression
    2. Genetics and Genomics
    Erik Toraason, Alina Salagean ... Diana E Libuda
    Research Article Updated

    The preservation of genome integrity during sperm and egg development is vital for reproductive success. During meiosis, the tumor suppressor BRCA1/BRC-1 and structural maintenance of chromosomes 5/6 (SMC-5/6) complex genetically interact to promote high fidelity DNA double strand break (DSB) repair, but the specific DSB repair outcomes these proteins regulate remain unknown. Using genetic and cytological methods to monitor resolution of DSBs with different repair partners in Caenorhabditis elegans, we demonstrate that both BRC-1 and SMC-5 repress intersister crossover recombination events. Sequencing analysis of conversion tracts from homolog-independent DSB repair events further indicates that BRC-1 regulates intersister/intrachromatid noncrossover conversion tract length. Moreover, we find that BRC-1 specifically inhibits error prone repair of DSBs induced at mid-pachytene. Finally, we reveal functional interactions of BRC-1 and SMC-5/6 in regulating repair pathway engagement: BRC-1 is required for localization of recombinase proteins to DSBs in smc-5 mutants and enhances DSB repair defects in smc-5 mutants by repressing theta-mediated end joining (TMEJ). These results are consistent with a model in which some functions of BRC-1 act upstream of SMC-5/6 to promote recombination and inhibit error-prone DSB repair, while SMC-5/6 acts downstream of BRC-1 to regulate the formation or resolution of recombination intermediates. Taken together, our study illuminates the coordinated interplay of BRC-1 and SMC-5/6 to regulate DSB repair outcomes in the germline.

    1. Chromosomes and Gene Expression
    Ryo Kariyazono, Takashi Osanai
    Research Article

    The hox operon in Synechocystis sp. PCC 6803, encoding bidirectional hydrogenase responsible for H2 production, is transcriptionally upregulated under microoxic conditions. Although several regulators for hox transcription have been identified, their dynamics and higher-order DNA structure of hox region in microoxic conditions remain elusive. We focused on key regulators for the hox operon: cyAbrB2, a conserved regulator in cyanobacteria, and SigE, an alternative sigma factor. Chromatin immunoprecipitation sequencing revealed that cyAbrB2 binds to the hox promoter region under aerobic conditions, with its binding being flattened in microoxic conditions. Concurrently, SigE exhibited increased localization to the hox promoter under microoxic conditions. Genome-wide analysis revealed that cyAbrB2 binds broadly to AT-rich genome regions and represses gene expression. Moreover, we demonstrated the physical interactions of the hox promoter region with its distal genomic loci. Both the transition to microoxic conditions and the absence of cyAbrB2 influenced the chromosomal interaction. From these results, we propose that cyAbrB2 is a cyanobacterial nucleoid-associated protein (NAP), modulating chromosomal conformation, which blocks RNA polymerase from the hox promoter in aerobic conditions. We further infer that cyAbrB2, with altered localization pattern upon microoxic conditions, modifies chromosomal conformation in microoxic conditions, which allows SigE-containing RNA polymerase to access the hox promoter. The coordinated actions of this NAP and the alternative sigma factor are crucial for the proper hox expression in microoxic conditions. Our results highlight the impact of cyanobacterial chromosome conformation and NAPs on transcription, which have been insufficiently investigated.