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Main conclusion <p><b>The 5′ UTRs of </b><Emphasis Type="BoldItalic">PEPCs</Emphasis><b> in</b><Emphasis Type="BoldItalic"> Suaeda aralocaspica</Emphasis><b> repress gene expression via intron sequence and upstream ORFs, and contain candidate </b><Emphasis Type="BoldItalic">cis</Emphasis><b>-acting elements that may be associated with differential transcriptional responses under ionic and osmotic stress conditions.</b></p> Abstract <p>The 5′ untranslated region (5′ UTR) plays a pivotal role in controlling gene expression during plant development and stress responses. It contains regulatory elements such as internal ribosome entry sites, upstream open reading frames (uORF), introns, and secondary structures, which collectively enable precisely translational control. Variations in these elements can give rise to multiple mRNA isoforms, thereby influencing gene expression levels. In this study, we analysed the 5′ UTR of two key photosynthetic enzyme genes, <i>PEPC1</i> and <i>PEPC2</i>, from <i>Suaeda aralocaspica</i>, a halophyte with C<sub>4</sub> photosynthesis pathway in a single polarised cell. GUS and dual-luciferase reporter assays examined the effects of mutation in the 5′ UTR, in particular its intron and uORFs on reporter gene expression. Both 5′ UTRs significantly repressed <i>PEPC</i> gene expression, with the <i>SaPEPC2</i> 5′ UTR exhibiting a stronger effect. Further dissection revealed that the <i>SaPEPC2</i> 5′ UTR harbours greater sequence and structural complexity, including one intron and four uORFs critical for its repressive function. Moreover, transgenic <i>Arabidopsis</i> carrying these 5′ UTRs showed differential reporter activity under NaCl and mannitol treatments, suggesting that candidate <i>cis</i>-acting elements within the 5′ UTR may be involved in abiotic stress-related regulation. These findings indicate the 5′ UTR as an important regulatory element for <i>PEPC</i> expression in heterologous reporter systems and provide a foundation for investigating its physiological role in halophytes.</p>

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Functional characterisation of the 5′ UTR of two PEPCs in Suaeda aralocaspica, a halophyte with C4 photosynthesis pathway in a single polarised cell

  • 期刊论文

  • 发表日期：

    2026年5月26日

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  • Zhimin Xu,
  • Yanxia Liu,
  • Mengyu Yan,
  • Haiyan Lan

摘要

Main conclusion

The 5′ UTRs of PEPCs in Suaeda aralocaspica repress gene expression via intron sequence and upstream ORFs, and contain candidate cis-acting elements that may be associated with differential transcriptional responses under ionic and osmotic stress conditions.

Abstract

The 5′ untranslated region (5′ UTR) plays a pivotal role in controlling gene expression during plant development and stress responses. It contains regulatory elements such as internal ribosome entry sites, upstream open reading frames (uORF), introns, and secondary structures, which collectively enable precisely translational control. Variations in these elements can give rise to multiple mRNA isoforms, thereby influencing gene expression levels. In this study, we analysed the 5′ UTR of two key photosynthetic enzyme genes, PEPC1 and PEPC2, from Suaeda aralocaspica, a halophyte with C4 photosynthesis pathway in a single polarised cell. GUS and dual-luciferase reporter assays examined the effects of mutation in the 5′ UTR, in particular its intron and uORFs on reporter gene expression. Both 5′ UTRs significantly repressed PEPC gene expression, with the SaPEPC2 5′ UTR exhibiting a stronger effect. Further dissection revealed that the SaPEPC2 5′ UTR harbours greater sequence and structural complexity, including one intron and four uORFs critical for its repressive function. Moreover, transgenic Arabidopsis carrying these 5′ UTRs showed differential reporter activity under NaCl and mannitol treatments, suggesting that candidate cis-acting elements within the 5′ UTR may be involved in abiotic stress-related regulation. These findings indicate the 5′ UTR as an important regulatory element for PEPC expression in heterologous reporter systems and provide a foundation for investigating its physiological role in halophytes.