Faculty of Biology, Technion - Israel Institute of Technology, Haifa, 32000, Israel.
Sci Rep. 2017 Jun 8;7(1):3037. doi: 10.1038/s41598-017-02873-z.
Members of the yeast family of PUF proteins bind unique subsets of mRNA targets that encode proteins with common functions. They therefore became a paradigm for post-transcriptional gene control. To provide new insights into the roles of the seemingly redundant Puf1 and Puf2 members, we monitored the growth rates of their deletions under many different stress conditions. A differential effect was observed at high CaCl concentrations, whereby puf1Δ growth was affected much more than puf2Δ, and inhibition was exacerbated in puf1Δpuf2Δ double knockout. Transcriptome analyses upon CaCl application for short and long terms defined the transcriptional response to CaCl and revealed distinct expression changes for the deletions. Intriguingly, mRNAs known to be bound by Puf1 or Puf2 were affected mainly in the double knockout. We focused on the cell wall regulator Zeo1 and observed that puf1Δpuf2Δ fails to maintain low levels of its mRNA. Complementarily, puf1Δpuf2Δ growth defect in CaCl was repaired upon further deletion of the Zeo1 gene. Thus, these proteins probably regulate the cell-wall integrity pathway by regulating Zeo1 post-transcriptionally. This work sheds new light on the roles of Puf proteins during the cellular response to environmental stress.
酵母家族的 PUF 蛋白成员结合独特的 mRNA 靶标子集，这些靶标编码具有共同功能的蛋白质。因此，它们成为转录后基因调控的范例。为了深入了解看似冗余的 Puf1 和 Puf2 成员的作用，我们在许多不同的应激条件下监测了它们缺失体的生长速率。在高 CaCl 浓度下观察到了不同的影响，puf1Δ 的生长受到的影响比 puf2Δ 大得多，并且在 puf1Δpuf2Δ 双敲除中抑制作用加剧。CaCl 短期和长期应用后的转录组分析定义了对 CaCl 的转录反应，并揭示了缺失体的明显表达变化。有趣的是，已知与 Puf1 或 Puf2 结合的 mRNA 主要在双敲除中受到影响。我们专注于细胞壁调节剂 Zeo1，并观察到 puf1Δpuf2Δ 无法维持其 mRNA 的低水平。补充的是，进一步缺失 Zeo1 基因修复了 puf1Δpuf2Δ 在 CaCl 中的生长缺陷。因此，这些蛋白质可能通过调节 Zeo1 的转录后水平来调节细胞壁完整性途径。这项工作为 Puf 蛋白在细胞对环境应激的反应中的作用提供了新的见解。
