A Protein Complex Required for Polymerase V Transcripts and RNA- Directed DNA Methylation in Arabidopsis
Type
ArticleAuthors
Law, Julie A.Ausín, Israel
Johnson, Lianna M.
Vashisht, Ajay A Amar
Zhu, Jian-Kang

Wohlschlegel, James A A.
Jacobsen, Steven E.
KAUST Department
Desert Agriculture InitiativeBiological and Environmental Sciences and Engineering (BESE) Division
Date
2010-04-23Embargo End Date
2011-04-23Permanent link to this record
http://hdl.handle.net/10754/561476
Metadata
Show full item recordAbstract
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.2010.03.062Sponsors
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 BVJournal
Current BiologyPubMed ID
20409711PubMed Central ID
PMC2972704Additional Links
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704
ae974a485f413a2113503eed53cd6c53
10.1016/j.cub.2010.03.062
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