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

Handle URI:
http://hdl.handle.net/10754/561476
Title:
A Protein Complex Required for Polymerase V Transcripts and RNA- Directed DNA Methylation in Arabidopsis
Authors:
Law, Julie A.; Ausín, Israel; Johnson, Lianna M.; Vashisht, Ajay A Amar; Zhu, Jian-Kang; Wohlschlegel, James A A.; Jacobsen, Steven E.
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.
KAUST Department:
Center for Desert Agriculture
Publisher:
Elsevier BV
Journal:
Current Biology
Issue Date:
May-2010
DOI:
10.1016/j.cub.2010.03.062
PubMed ID:
20409711
PubMed Central ID:
PMC2972704
Type:
Article
ISSN:
09609822
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.
Additional Links:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704
Appears in Collections:
Articles; Center for Desert Agriculture

Full metadata record

DC FieldValue Language
dc.contributor.authorLaw, Julie A.en
dc.contributor.authorAusín, Israelen
dc.contributor.authorJohnson, Lianna M.en
dc.contributor.authorVashisht, Ajay A Amaren
dc.contributor.authorZhu, Jian-Kangen
dc.contributor.authorWohlschlegel, James A A.en
dc.contributor.authorJacobsen, Steven E.en
dc.date.accessioned2015-08-02T09:12:20Zen
dc.date.available2015-08-02T09:12:20Zen
dc.date.issued2010-05en
dc.identifier.issn09609822en
dc.identifier.pmid20409711en
dc.identifier.doi10.1016/j.cub.2010.03.062en
dc.identifier.urihttp://hdl.handle.net/10754/561476en
dc.description.abstractDNA 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.en
dc.description.sponsorshipWe 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.en
dc.publisherElsevier BVen
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2972704en
dc.subjectDNAen
dc.subjectRNAen
dc.titleA Protein Complex Required for Polymerase V Transcripts and RNA- Directed DNA Methylation in Arabidopsisen
dc.typeArticleen
dc.contributor.departmentCenter for Desert Agricultureen
dc.identifier.journalCurrent Biologyen
dc.identifier.pmcidPMC2972704en
dc.contributor.institutionDepartment of Molecular Cell and Developmental Biology, University of California at Los Angeles, Los Angeles, CA 90095, United Statesen
dc.contributor.institutionLife Sciences Core Curriculum, University of California Los Angeles, Los Angeles, CA 90095, United Statesen
dc.contributor.institutionDepartment of Botany and Plant Sciences, University of California, Riverside, Riverside, CA 92521, United Statesen
dc.contributor.institutionDepartment of Biological Chemistry, David Geffen School of Medicine, UCLA, BSRB-377A, 615 Charles E. Young Dr. South, Los Angeles, CA 90095-1737, United Statesen
dc.contributor.institutionHoward Hughes Medical Institute, University of California at Los Angeles, Los Angeles, CA 90095, United Statesen
kaust.authorZhu, Jian-Kangen

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