Transgene-induced silencing of Arabidopsis phytochrome A gene via exonic methylation

被引:20
作者
Chawla, Rekha
Nicholson, Scott J.
Folta, Kevin M.
Srivastava, Vibha [1 ]
机构
[1] Univ Arkansas, Dept Crop & Soil Environm Sci, Fayetteville, AR 72701 USA
[2] Univ Florida, Dept Hort Sci, Gainesville, FL 32611 USA
[3] Univ Arkansas, Dept Hort, Fayetteville, AR 72701 USA
关键词
epi-allele; DNA methylation; chromatin modification; transcriptional silencing; phytochrome A;
D O I
10.1111/j.1365-313X.2007.03301.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Transgene-induced promoter or enhancer methylation clearly retards gene activity. While exonic methylation of genes is frequently observed in the RNAi process, only sporadic evidence has demonstrated its definitive role in gene suppression. Here, we report the isolation of a transcriptionally suppressed epi-allele of the Arabidopsis thaliana phytochrome A gene (PHYA) termed phyA' that shows methylation only in symmetric CG sites resident in exonic regions. These exonic modifications confer a strong phyA mutant phenotype, characterized by elongated hypocotyls in seedlings grown under continuous far-red light. De-methylation of phyA' in the DNA methyl transferase I (met1) mutant background increased PHYA expression and restored the wild-type phenotype, confirming the pivotal role of exonic CG methylation in maintaining the altered epigenetic state. PHYA epimutation was apparently induced by a transgene locus; however, it is stably maintained following segregation. Chromatin immunoprecipitation assays revealed association with dimethyl histone H3 lysine 9 (H3K9me2), a heterochromatic marker, within the phyA' coding region. Therefore, transgene-induced exonic methylation can lead to chromatin alteration that affects gene expression, most likely through reduction in the transcription rate.
引用
收藏
页码:1105 / 1118
页数:14
相关论文
共 71 条
[21]   Plant DNA methyltransferases [J].
Finnegan, EJ ;
Kovac, KA .
PLANT MOLECULAR BIOLOGY, 2000, 43 (2-3) :189-201
[22]   Epigenetic switch from posttranscriptional to transcriptional silencing is correlated with promoter hypermethylation [J].
Fojtova, M ;
Van Houdt, H ;
Depicker, A ;
Kovarik, A .
PLANT PHYSIOLOGY, 2003, 133 (03) :1240-1250
[23]   Isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays [J].
Folta, Kevin M. ;
Kaufman, Lon S. .
NATURE PROTOCOLS, 2006, 1 (06) :3094-3100
[24]   Preparation of transcriptionally active nuclei from etiolated Arabidopsis thaliana [J].
Folta, KM ;
Kaufman, LS .
PLANT CELL REPORTS, 2000, 19 (05) :504-510
[25]   Alternative silencing effects involve distinct types of non-spreading cytosine methylation at a three-gene, single-copy transgenic locus in rice [J].
Fu, X ;
Kohli, A ;
Twyman, RM ;
Christou, P .
MOLECULAR AND GENERAL GENETICS, 2000, 263 (01) :106-118
[26]   Transcriptional silencing by RNAi in the soma of of a transgene C. elegans [J].
Grishok, A ;
Sinskey, JL ;
Sharp, PA .
GENES & DEVELOPMENT, 2005, 19 (06) :683-696
[27]   Conditional circadian regulation of PHYTOCHROME A gene expression [J].
Hall, A ;
Kozma-Bognár, L ;
Tóth, R ;
Nagy, F ;
Millar, AJ .
PLANT PHYSIOLOGY, 2001, 127 (04) :1808-1818
[28]   Identification and characterization of a family of mammalian methyl-CpG binding proteins [J].
Hendrich, B ;
Bird, A .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (11) :6538-6547
[29]   Methylation of coding region alone inhibits gene expression in plant protoplasts [J].
Hohn, T ;
Corsten, S ;
Rieke, S ;
Muller, M ;
Rothnie, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (16) :8334-8339
[30]   Stability of patch methylation and its impact in regions of transcriptional initiation and elongation [J].
Hsieh, CL .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (10) :5897-5904