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    A Protein Complex Required for Polymerase V Transcripts and RNA- Directed DNA Methylation in Arabidopsis

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    Type
    Article
    Authors
    Law, Julie A.
    Ausín, Israel
    Johnson, Lianna M.
    Vashisht, Ajay A Amar
    Zhu, Jian-Kang cc
    Wohlschlegel, James A A.
    Jacobsen, Steven E.
    KAUST Department
    Desert Agriculture Initiative
    Biological and Environmental Sciences and Engineering (BESE) Division
    Date
    2010-04-23
    Embargo End Date
    2011-04-23
    Permanent link to this record
    http://hdl.handle.net/10754/561476
    
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    Abstract
    DNA methylation is an epigenetic modification associated with gene silencing. In Arabidopsis, DNA methylation is established by DOMAINS REARRANGED METHYLTRANSFERASE 2 (DRM2), which is targeted by small interfering RNAs through a pathway termed RNA-directed DNA methylation (RdDM) [1, 2]. Recently, RdDM was shown to require intergenic noncoding (IGN) transcripts that are dependent on the Pol V polymerase. These transcripts are proposed to function as scaffolds for the recruitment of downstream RdDM proteins, including DRM2, to loci that produce both siRNAs and IGN transcripts [3]. However, the mechanism(s) through which Pol V is targeted to specific genomic loci remains largely unknown. Through affinity purification of two known RdDM components, DEFECTIVE IN RNA-DIRECTED DNA METHYLATION 1 (DRD1) [4] and DEFECTIVE IN MERISTEM SILENCING 3 (DMS3) [5, 6], we found that they copurify with each other and with a novel protein, RNA-DIRECTED DNA METHYLATION 1 (RDM1), forming a complex we term DDR. We also found that DRD1 copurified with Pol V subunits and that RDM1, like DRD1 [3] and DMS3 [7], is required for the production of Pol V-dependent transcripts. These results suggest that the DDR complex acts in RdDM at a step upstream of the recruitment or activation of Pol V. © 2010 Elsevier Ltd. All rights reserved.
    Citation
    Law, J. A., Ausin, I., Johnson, L. M., Vashisht, A. A., Zhu, J.-K., Wohlschlegel, J. A., & Jacobsen, S. E. (2010). A Protein Complex Required for Polymerase V Transcripts and RNA- Directed DNA Methylation in Arabidopsis. Current Biology, 20(10), 951–956. doi:10.1016/j.cub.2010.03.062
    Sponsors
    We thank T. LaGrange for providing the NRPE1 antibody and members of the Jacobsen laboratory for helpful discussion. Jacobsen lab research was supported by U.S. National Institutes of Health (NIH) grant GM60398. I.A. was supported by a postdoctoral fellowship from the Ministerio de Educacion y Ciencia. J.A.L. was supported the NIH National Research Service Award 5F32GM820453. Wohlschlegel lab research was supported by University of California, Los Angeles Jonsson Cancer Center. S.E.J. is an investigator of the Howard Hughes Medical Institute. S.E.J., JAW., J.A.L., and I.A designed the experiments; J.A.L, I.A., L.M.J, and A.A.V. performed the experiments; J.K.Z. provided the ros1-1 rdm1-1 mutant allele and the RDM1 antibody; J.A.L. wrote the paper.
    Publisher
    Elsevier BV
    Journal
    Current Biology
    DOI
    10.1016/j.cub.2010.03.062
    PubMed ID
    20409711
    PubMed Central ID
    PMC2972704
    Additional Links
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.cub.2010.03.062
    Scopus Count
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    Articles; Biological and Environmental Sciences and Engineering (BESE) Division; Desert Agriculture Initiative

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