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Published on: April 19, 2011
Cabbage Peptide miPEP156a Enhances the Level of Accumulation of Its mRNA in Transgenic Moss Physcomitrium patens
T N Erokhina1, E V Ryabukhina1, I S Lyapina1
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997 Russia.
Abstract:
MicroRNAs are endogenous, small non-coding RNAs that regulate gene expression at the post-transcriptional level by cleaving target mRNAs. Mature microRNAs are products of the processing of their primary transcripts (pri-miRNAs). Now, it has been discovered that the products of the translation of some plant pri-miRNAs are peptide molecules (miPEP). These peptides have the capacity to physically interact with their open reading frames (ORFs) in the transcribed pri-miRNAs and, thus, positively regulate the accumulation of these RNAs and the corresponding mature microRNAs. Most conserved microRNAs play an important role in plants development and their response to stress. In this work, we obtained transgenic Physcomitrium patens moss plants containing Brassica oleracea miPEP156a ORF in the genome under the control of a strong 35S cauliflower mosaic virus promoter and analyzed the effect of the exogenous peptide on the transcription of this ORF in the protonemata of two transgenic moss lines. It turned out that the chemically synthesized peptide miPEP156a increases the accumulation of its own mRNA during moss culture growth, as was previously shown in studies by foreign researchers and in our own work for a number of peptides in monocotyledonous and dicotyledonous plants. These findings confirm that pri-miRNA regions that are located outside the coding region of the peptide are not required for transcriptional activation. Moreover, we have also succeeded in showing that the presence of a specific promoter of the microRNA gene does not affect the phenomenon of transcription activation; this phenomenon per se is not species-specific and is observed in transgenic plants, regardless of the origin of the miPEP.
Insights
Plant micropeptides (miPEPs) can enhance their own RNA accumulation, demonstrating a conserved, species-independent mechanism. This discovery reveals a novel layer of gene regulation in plants, impacting development and stress responses.
Area of Science:
- Molecular Biology
- Plant Science
- Gene Regulation
Background:
- MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression post-transcriptionally.
- Plant primary miRNA transcripts (pri-miRNAs) can be translated into micropeptides (miPEPs).
- miPEPs can interact with their own open reading frames (ORFs) to enhance pri-miRNA accumulation.
Purpose of the Study:
- To investigate the effect of an exogenous Brassica oleracea miPEP156a peptide on its own mRNA transcription in transgenic Physcomitrium patens moss.
- To determine if pri-miRNA regions outside the miPEP ORF are necessary for transcriptional activation.
- To assess the species-specificity and promoter-dependence of the miPEP-mediated transcriptional activation phenomenon.
Main Methods:
- Generation of transgenic Physcomitrium patens moss expressing Brassica oleracea miPEP156a ORF under a 35S promoter.
- Analysis of miPEP156a mRNA accumulation in protonemata of transgenic moss lines.
- Chemical synthesis of miPEP156a peptide for experimental validation.
Main Results:
- Chemically synthesized miPEP156a peptide increased the accumulation of its own mRNA in moss protonemata.
- Transcriptional activation of pri-miRNA occurred independently of regions outside the miPEP coding sequence.
- The phenomenon of transcriptional activation by miPEPs is not species-specific and occurs regardless of the specific miRNA gene promoter.
Conclusions:
- Plant miPEPs can autoregulate their own transcript levels, confirming a conserved regulatory mechanism.
- The miPEP-mediated transcriptional activation is independent of the pri-miRNA sequence flanking the ORF and the specific promoter used.
- This autoregulatory mechanism is conserved across plant species, offering a novel target for modulating miRNA levels and plant traits.
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