Sense transgene-induced post-transcriptional gene silencing in tobacco compromises the splicing of endogenous counterpart genes.
Sense transgene-induced post-transcriptional gene silencing in tobacco compromises the splicing of endogenous counterpart genes.
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DOI：
10.1371/journal.pone.0087869
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Kodama H
中科院分区：
文献类型：
--
作者：
Shin MR;Natsuume M;Matsumoto T;Hanaoka M;Imai M;Iijima K;Oka S;Adachi E;Kodama H
正义转基因诱导转录后基因沉默被认为是一种RNA沉默，在这种沉默中，ARGONAUTE1指导小干扰RNA(SiRNA)介导的靶基因在细胞质中的切割。在这里，我们报告了内源对应基因的剪接变化是其成熟mRNA水平降低的主要原因。烟草内质网ω-3脂肪酸脱氢酶(NtFAD3)基因的S-pTGS后，在总RNA组分中检测到3‘-截短的、多腺化的内切的3’-FAD3转录本。虽然转录一直持续到内源NtFAD3基因的最后一个外显子，但含有内含子的NtFAD3转录本在S-pTGS植株的细胞核中积累。当内源沉默抑制基因rgs-CaM在S-pTGS植株中过表达时，产生了几个含有大部分外显子序列的含有内含子的NtFAD3转录本。这些含有内含子的NtFAD3剪接变体是在存在与这些剪接变体的核苷酸序列同源的NtFAD3 siRNAs的情况下产生的。本研究的结果表明，抑制Endo-NtFAD3基因的表达主要是通过剪接的改变而不是通过细胞质切片器的活性。我们的结果表明，转基因和含有内含子的内源对应基因在S-pTGS植株中受到不同程度的抑制。
Sense transgene-induced post-transcriptional gene silencing (S-PTGS) is thought to be a type of RNA silencing in which ARGONAUTE1 directs the small interfering RNA (siRNA)-mediated cleavage of a target mRNA in the cytoplasm. Here, we report that the altered splicing of endogenous counterpart genes is a main cause for the reduction of their mature mRNA levels. After the S-PTGS of a tobacco endoplasmic reticulum ω-3 fatty acid desaturase (NtFAD3) gene, 3′-truncated, polyadenylated endo-NtFAD3 transcripts and 5′-truncated, intron-containing endo-NtFAD3 transcripts were detected in the total RNA fraction. Although transcription proceeded until the last exon of the endogenous NtFAD3 gene, intron-containing NtFAD3 transcripts accumulated in the nucleus of the S-PTGS plants. Several intron-containing NtFAD3 transcripts harboring most of the exon sequences were generated when an endogenous silencing suppressor gene, rgs-CaM, was overexpressed in the S-PTGS plants. These intron-containing NtFAD3 splice variants were generated in the presence of NtFAD3 siRNAs that are homologous to the nucleotide sequences of these splice variants. The results of this study indicate that the inhibition of endo-NtFAD3 gene expression is primarily directed via the alteration of splicing and not by cytoplasmic slicer activity. Our results suggest that the transgene and intron-containing endogenous counterpart genes are differentially suppressed in S-PTGS plants.
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影响因子：
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作者：
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