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Post-transcriptional silencing of a neomycin phosphotransferase II transgene correlates with the accumulation of unproductive RNAs and with increased cytosine methylation of 3' flanking regions

Helena Van Houdt (UGent) , Ivan Ingelbrecht (UGent) , Marc Van Montagu (UGent) and Anna Depicker (UGent)
(1997) PLANT JOURNAL. 12(2). p.379-392
Author
Organization
Abstract
The transgenic tobacco plant GVCHS(320)-1 harbours several T-DNAs with the neomycin phosphotransferase II (nptII)-encoding chimeric gene under control of the cauliflower mosaic virus 35S promoter (CaMV 35S). These T-DNAs are distributed over two loci, named A and B. The primary transformant GVCHS(320)-1 had substantially reduced steady state nptII transcript levels due to the activity of a post transcriptional silencing mechanism. Silencing of the nptII-encoding transgenes in the progeny of GVCHS(320)-1 requires the presence of the A locus that consists of two physically separated T-DNAs. Although the B locus contains three T-DNAs in the same orientation, it gives rise to high steady-state nptII mRNA levels and NPTII protein levels even in homozygous condition. The B locus only becomes silenced in trans in the presence of a silencing locus. The data suggest that silencing of the nptII transgenes is reinforced by ageing. It is also suggested that silencing is correlated with a reduced NPTII protein/nptII mRNA ratio, which may be interpreted as the accumulation of unproductive nptII RNA molecules in silenced plants. The data further demonstrate that the silenced state is correlated with extensive C-methylation of diagnostic sites in the 3' three-quarters of the coding sequence and in the 3' region up to 1400 bp downstream of the polyadenylation signal.
Keywords
LEVEL, TOBACCO, GENE-EXPRESSION, PETUNIA-HYBRIDA, SUPPRESSION, LOCI, VIRUS-RESISTANCE, DNA-METHYLATION, INACTIVATION, PLANTS

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Citation

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MLA
Van Houdt, Helena, Ivan Ingelbrecht, Marc Van Montagu, et al. “Post-transcriptional Silencing of a Neomycin Phosphotransferase II Transgene Correlates with the Accumulation of Unproductive RNAs and with Increased Cytosine Methylation of 3’ Flanking Regions.” PLANT JOURNAL 12.2 (1997): 379–392. Print.
APA
Van Houdt, H., Ingelbrecht, I., Van Montagu, M., & Depicker, A. (1997). Post-transcriptional silencing of a neomycin phosphotransferase II transgene correlates with the accumulation of unproductive RNAs and with increased cytosine methylation of 3’ flanking regions. PLANT JOURNAL, 12(2), 379–392.
Chicago author-date
Van Houdt, Helena, Ivan Ingelbrecht, Marc Van Montagu, and Anna Depicker. 1997. “Post-transcriptional Silencing of a Neomycin Phosphotransferase II Transgene Correlates with the Accumulation of Unproductive RNAs and with Increased Cytosine Methylation of 3’ Flanking Regions.” Plant Journal 12 (2): 379–392.
Chicago author-date (all authors)
Van Houdt, Helena, Ivan Ingelbrecht, Marc Van Montagu, and Anna Depicker. 1997. “Post-transcriptional Silencing of a Neomycin Phosphotransferase II Transgene Correlates with the Accumulation of Unproductive RNAs and with Increased Cytosine Methylation of 3’ Flanking Regions.” Plant Journal 12 (2): 379–392.
Vancouver
1.
Van Houdt H, Ingelbrecht I, Van Montagu M, Depicker A. Post-transcriptional silencing of a neomycin phosphotransferase II transgene correlates with the accumulation of unproductive RNAs and with increased cytosine methylation of 3’ flanking regions. PLANT JOURNAL. 1997;12(2):379–92.
IEEE
[1]
H. Van Houdt, I. Ingelbrecht, M. Van Montagu, and A. Depicker, “Post-transcriptional silencing of a neomycin phosphotransferase II transgene correlates with the accumulation of unproductive RNAs and with increased cytosine methylation of 3’ flanking regions,” PLANT JOURNAL, vol. 12, no. 2, pp. 379–392, 1997.
@article{185010,
  abstract     = {The transgenic tobacco plant GVCHS(320)-1 harbours several T-DNAs with the neomycin phosphotransferase II (nptII)-encoding chimeric gene under control of the cauliflower mosaic virus 35S promoter (CaMV 35S). These T-DNAs are distributed over two loci, named A and B. The primary transformant GVCHS(320)-1 had substantially reduced steady state nptII transcript levels due to the activity of a post transcriptional silencing mechanism. Silencing of the nptII-encoding transgenes in the progeny of GVCHS(320)-1 requires the presence of the A locus that consists of two physically separated T-DNAs. Although the B locus contains three T-DNAs in the same orientation, it gives rise to high steady-state nptII mRNA levels and NPTII protein levels even in homozygous condition. The B locus only becomes silenced in trans in the presence of a silencing locus. The data suggest that silencing of the nptII transgenes is reinforced by ageing. It is also suggested that silencing is correlated with a reduced NPTII protein/nptII mRNA ratio, which may be interpreted as the accumulation of unproductive nptII RNA molecules in silenced plants. The data further demonstrate that the silenced state is correlated with extensive C-methylation of diagnostic sites in the 3' three-quarters of the coding sequence and in the 3' region up to 1400 bp downstream of the polyadenylation signal.},
  author       = {Van Houdt, Helena and Ingelbrecht, Ivan and Van Montagu, Marc and Depicker, Anna},
  issn         = {0960-7412},
  journal      = {PLANT JOURNAL},
  keywords     = {LEVEL,TOBACCO,GENE-EXPRESSION,PETUNIA-HYBRIDA,SUPPRESSION,LOCI,VIRUS-RESISTANCE,DNA-METHYLATION,INACTIVATION,PLANTS},
  language     = {eng},
  number       = {2},
  pages        = {379--392},
  title        = {Post-transcriptional silencing of a neomycin phosphotransferase II transgene correlates with the accumulation of unproductive RNAs and with increased cytosine methylation of 3' flanking regions},
  url          = {http://dx.doi.org/10.1046/j.1365-313X.1997.12020379.x},
  volume       = {12},
  year         = {1997},
}

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