Arabidopsis RPT2a, 19S proteasome subunit, regulates gene silencing via DNA methylation

The ubiquitin/proteasome pathway plays a crucial role in many biological processes. Here we report a novel role for the Arabidopsis 19S proteasome subunit RPT2a in regulating gene activity at the transcriptional level via DNA methylation. Knockout mutation of the RPT2a gene did not alter global prot...

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Bibliographic Details
Published in:PloS one 2012-05, Vol.7 (5), p.e37086-e37086
Main Authors: Sako, Kaori, Maki, Yuko, Kanai, Tomoyuki, Kato, Eriko, Maekawa, Shugo, Yasuda, Shigetaka, Sato, Takeo, Watahiki, Masaaki K, Yamaguchi, Junji
Format: Article
Language:English
Subjects:
Arabidopsis
Arabidopsis - genetics
Arabidopsis Proteins - genetics
Arabidopsis Proteins - metabolism
Arabidopsis thaliana
Biological activity
Biology
Bisulfite
Comparative analysis
Cytokines
Deoxyribonucleic acid
DNA
DNA Methylation
DNA methyltransferase
Epigenetics
Gene expression
Gene Expression Regulation, Plant - genetics
Gene Silencing
Genetic aspects
Genomes
Histone deacetylase
Histone Deacetylases - metabolism
Inhibitors
Methylation
Mutants
Mutation
Phenols
Promoter Regions, Genetic
Proteasome Endopeptidase Complex - genetics
Proteasomes
Proteins
RNA polymerase
Transcription (Genetics)
Transcription, Genetic
Transgenes
Transgenes - genetics
Transgenic plants
Transposons
Trichostatin A
Ubiquitin
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Summary:The ubiquitin/proteasome pathway plays a crucial role in many biological processes. Here we report a novel role for the Arabidopsis 19S proteasome subunit RPT2a in regulating gene activity at the transcriptional level via DNA methylation. Knockout mutation of the RPT2a gene did not alter global protein levels; however, the transcriptional activities of reporter transgenes were severely reduced compared to those in the wild type. This transcriptional gene silencing (TGS) was observed for transgenes under control of either the constitutive CaMV 35S promoter or the cold-inducible RD29A promoter. Bisulfite sequencing analysis revealed that both the transgene and endogenous RD29A promoter regions were hypermethylated at CG and non-CG contexts in the rpt2a mutant. Moreover, the TGS of transgenes driven by the CaMV 35S promoters was released by treatment with the DNA methylation inhibitor 5-aza-2'-deoxycytidine, but not by application of the inhibitor of histone deacetylase Trichostatin A. Genetic crosses with the DNA methyltransferase met1 single or drm1drm2cmt3 triple mutants also resulted in a release of CaMV 35S transgene TGS in the rpt2a mutant background. Increased methylation was also found at transposon sequences, suggesting that the 19S proteasome containing AtRPT2a negatively regulates TGS at transgenes and at specific endogenous genes through DNA methylation.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0037086