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<p>Small regulatory RNAs (sRNAs), including small interfering (si) and micro (mi) RNAs, are components of the RNA interference (RNAi) pathways regulating vital processes. These various sRNAs act via specialized executor ARGONAUTE (AGO) proteins constituting RNA induced silencing complexes (RISCs). In <i>Arabidopsis</i>, miRNAs load mainly into AGO1, while 24-nucleotide (nt) long siRNAs load into AGO4, typically. However, a large subset of sRNAs is present in the cytoplasm in AGO-unbound form. To uncover the potential importance of tissue specific AGO-loading efficiency of sRNAs in an economically important plant we used pepper fruits, dissected into pericarp, placenta, and seed and the ratio between AGO-loaded and -unloaded sRNA pools was investigated with high throughput sequencing. In agreement with the previous studies, variable loading capacity of sRNAs was discovered with an abundant AGO-unbound pool. The investigated pools showed a surprisingly high prevalence of 21-22-24-nt long siRNAs even in the high molecular weight RISC. The dynamically expanding pericarp exhibited extremely high 21-nt long miRNA content. Moreover, comparing the different tissues, a drastic redistribution or elimination of 21- and 24-nt sRNAs in various sRNA-protein complexes and changes in unbound sRNA pools were observed demonstrating the highly dynamic alteration in AGO-loading properties of the sRNAs. These data imply that during fruit development sRNAs are dynamically sorted into various executor complexes in a controlled manner. This finding also implies that analysis of the content of AGO-sRNA complexes together with the AGO-unbound population of these sRNAs can be a useful tool to identify potentially functional sRNAs.</p>

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Tissue specific RISC-loading reassesses small RNA functionality in developing pepper fruit

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月11日

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  • Ágnes Dalmadi,
  • Péter Gyula,
  • Auwalu Abdu,
  • András Kis,
  • Fabio Miloro,
  • Jeannette Bálint,
  • György Szittya,
  • Zoltán Havelda

摘要

Small regulatory RNAs (sRNAs), including small interfering (si) and micro (mi) RNAs, are components of the RNA interference (RNAi) pathways regulating vital processes. These various sRNAs act via specialized executor ARGONAUTE (AGO) proteins constituting RNA induced silencing complexes (RISCs). In Arabidopsis, miRNAs load mainly into AGO1, while 24-nucleotide (nt) long siRNAs load into AGO4, typically. However, a large subset of sRNAs is present in the cytoplasm in AGO-unbound form. To uncover the potential importance of tissue specific AGO-loading efficiency of sRNAs in an economically important plant we used pepper fruits, dissected into pericarp, placenta, and seed and the ratio between AGO-loaded and -unloaded sRNA pools was investigated with high throughput sequencing. In agreement with the previous studies, variable loading capacity of sRNAs was discovered with an abundant AGO-unbound pool. The investigated pools showed a surprisingly high prevalence of 21-22-24-nt long siRNAs even in the high molecular weight RISC. The dynamically expanding pericarp exhibited extremely high 21-nt long miRNA content. Moreover, comparing the different tissues, a drastic redistribution or elimination of 21- and 24-nt sRNAs in various sRNA-protein complexes and changes in unbound sRNA pools were observed demonstrating the highly dynamic alteration in AGO-loading properties of the sRNAs. These data imply that during fruit development sRNAs are dynamically sorted into various executor complexes in a controlled manner. This finding also implies that analysis of the content of AGO-sRNA complexes together with the AGO-unbound population of these sRNAs can be a useful tool to identify potentially functional sRNAs.