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Ribosome binding of phasiRNA precursors accelerates the 24-nt phasiRNA burst in meiotic maize anthers
The Plant Cell ( IF 10.0 ) Pub Date : 2024-10-23 , DOI: 10.1093/plcell/koae289
Yingjia Han, Siqi Jiang, Xiaomei Dong, Xing Dai, Shunxi Wang, Ying Zheng, Ge Yan, Shengben Li, Liuji Wu, Virginia Walbot, Blake C Meyers, Mei Zhang

Reproductive phasiRNAs (phased, small interfering RNAs), produced from numerous PHAS loci, are essential for plant anther development. PHAS transcripts are enriched on endoplasmic reticulum-bound ribosomes in maize (Zea mays), but the impact of ribosome binding on phasiRNA biogenesis remains elusive. Through ribosome profiling of maize anthers at 10 developmental stages, we demonstrated that 24-PHAS transcripts are bound by ribosomes, with patterns corresponding to the timing and abundance of 24-PHAS transcripts. Ribosome binding to 24-PHAS transcripts is conserved among different maize inbred lines, with ribosomes enriched upstream of miR2275 target sites. We detected short open reading frames (sORFs) in the ribosome-binding regions of some 24-PHAS transcripts and observed a 3-nt periodicity in most sORFs, but mass spectrometry failed to detect peptides corresponding to the sORFs. Deletion of the entire ribosome-binding region of 24PHAS_NO296 locus eliminated ribosome binding and decreased 24-nt phasiRNA production, without affecting 24PHAS_NO296 transcript levels. In contrast, disrupting only the sORFs in 24PHAS_NO296 did not substantially affect the generation of 24-nt phasiRNAs. A newly formed sORF in these mutants may have re-directed ribosome binding to its transcripts. Overall, these findings demonstrate that sORFs facilitate ribosome binding to 24-PHAS transcripts, thereby promoting phasiRNA biogenesis in meiotic anthers.

中文翻译:


phasiRNA 前体的核糖体结合加速减数分裂玉米花药中的 24-nt phasiRNA 爆发



由许多 PHAS 基因座产生的生殖 phasiRNA (phased, small 干扰 RNA) 对植物花药发育至关重要。PHAS 转录本在玉米 (Zea mays) 中内质网结合的核糖体上富集,但核糖体结合对 phasiRNA 生物发生的影响仍然难以捉摸。通过对 10 个发育阶段的玉米花药进行核糖体分析,我们证明 24-PHAS 转录物与核糖体结合,其模式对应于 24-PHAS 转录物的时间和丰度。核糖体与 24-PHAS 转录本的结合在不同的玉米自交系中是保守的,核糖体在 miR2275 靶位点的上游富集。我们在一些 24-PHAS 转录本的核糖体结合区检测到短开放阅读框 (sORF),并在大多数 sORF 中观察到 3-nt 周期性,但质谱法未能检测到对应于 sORFs 的肽。缺失 24PHAS_NO296 位点的整个核糖体结合区消除了核糖体结合并减少了 24-nt phasiRNA 的产生,而不会影响24PHAS_NO296转录水平。相比之下,仅破坏 24PHAS_NO296 中的 sORF 不会对 24-nt phasiRNA 的产生产生实质性影响。这些突变体中新形成的 sORF 可能已将核糖体结合重定向到其转录本。总体而言,这些发现表明 sORFs 促进核糖体与 24-PHAS 转录本的结合,从而促进减数分裂花药中的 phasiRNA 生物发生。
更新日期:2024-10-23
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