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<p>As a gasotransmitter, hydrogen sulfide (H<sub>2</sub>S) predominantly modulates physiological processes through protein persulfidation. RBM25 is a splicing factor involved in alternative splicing. Nevertheless, it remains unclear whether H<sub>2</sub>S can extensively regulate the <i>Arabidopsis thaliana</i> genome via persulfidation of AtRBM25. Our results revealed that H<sub>2</sub>S persulfidates AtRBM25 at Cys226. <i>RNA-seq</i> showed a substantial genome-wide response to H<sub>2</sub>S, affecting both transcriptional and post-transcriptional levels. GO enrichment analysis indicated that H<sub>2</sub>S-regulated genes were significantly enriched in stress‑response terms. Among the AS events induced by H<sub>2</sub>S in WT, the A3SS became the most prevalent type (34.37%), followed by RI, A5SS and SE. The transcriptional response to H<sub>2</sub>S partially relied on AtRBM25, and the differentially expressed genes were enriched in GO terms associated with abiotic stress. RT-qPCR confirmed that H<sub>2</sub>S-mediated regulation of the stress-responsive genes <i>GRXS13</i>, <i>ARR4</i>, <i>HAI1</i>, <i>GSTF7</i>, <i>AFP3</i> and <i>FBA5</i> was dependent on AtRBM25. Notably, this H<sub>2</sub>S-induced AS profile was partially dependent on AtRBM25, as the proportion of A3SS further increased to 39.27%, whereas other event types showed minimal changes. GO and KEGG analyses of these AS genes showed significant enrichment in RNA splicing pathways. Specifically, the responsiveness of different transcript isoforms of <i>HAI1</i>, <i>AFP3</i>, and <i>ARR4</i> to H<sub>2</sub>S varied in their dependence on AtRBM25. Collectively, H<sub>2</sub>S-mediated persulfidation of AtRBM25 not only regulates downstream gene expression, but also participates in splicing reprogramming. Taken together, these findings provide pivotal clues into H<sub>2</sub>S-regulated alternative splicing <i>via</i> AtRBM25, and reveal a novel perspective of H<sub>2</sub>S-mediated transcriptional and post-transcriptional coordination through post-translational modification.</p>

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RNA-seq Reveals Genome-Wide H2S Modulation of Transcription and Splicing Through AtRBM25 Persulfidation in Arabidopsis

  • 期刊论文

  • 发表日期：

    2026年6月22日

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  • Zhangjing Wu,
  • Liping Zhang,
  • Saisai Pei,
  • Wenqi Liu,
  • Hua Li,
  • Yanxi Pei

摘要

As a gasotransmitter, hydrogen sulfide (H2S) predominantly modulates physiological processes through protein persulfidation. RBM25 is a splicing factor involved in alternative splicing. Nevertheless, it remains unclear whether H2S can extensively regulate the Arabidopsis thaliana genome via persulfidation of AtRBM25. Our results revealed that H2S persulfidates AtRBM25 at Cys226. RNA-seq showed a substantial genome-wide response to H2S, affecting both transcriptional and post-transcriptional levels. GO enrichment analysis indicated that H2S-regulated genes were significantly enriched in stress‑response terms. Among the AS events induced by H2S in WT, the A3SS became the most prevalent type (34.37%), followed by RI, A5SS and SE. The transcriptional response to H2S partially relied on AtRBM25, and the differentially expressed genes were enriched in GO terms associated with abiotic stress. RT-qPCR confirmed that H2S-mediated regulation of the stress-responsive genes GRXS13, ARR4, HAI1, GSTF7, AFP3 and FBA5 was dependent on AtRBM25. Notably, this H2S-induced AS profile was partially dependent on AtRBM25, as the proportion of A3SS further increased to 39.27%, whereas other event types showed minimal changes. GO and KEGG analyses of these AS genes showed significant enrichment in RNA splicing pathways. Specifically, the responsiveness of different transcript isoforms of HAI1, AFP3, and ARR4 to H2S varied in their dependence on AtRBM25. Collectively, H2S-mediated persulfidation of AtRBM25 not only regulates downstream gene expression, but also participates in splicing reprogramming. Taken together, these findings provide pivotal clues into H2S-regulated alternative splicing via AtRBM25, and reveal a novel perspective of H2S-mediated transcriptional and post-transcriptional coordination through post-translational modification.