Plant Developmental Genetics, Institute of Biology Leiden, Leiden University, Leiden, the Netherlands.
Plant Systems Biology, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany.
Nat Plants. 2020 May;6(5):544-555. doi: 10.1038/s41477-020-0650-2. Epub 2020 May 11.
The 3-phosphoinositide-dependent protein kinase 1 (PDK1) is a conserved master regulator of AGC kinases in eukaryotic organisms. pdk1 loss of function causes a lethal phenotype in animals and yeasts, but only mild phenotypic defects in Arabidopsis thaliana (Arabidopsis). The Arabidopsis genome contains two PDK1-encoding genes, PDK1 and PDK2. Here, we used clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) to generate true loss-of-function pdk1 alleles, which, when combined with pdk2 alleles, showed severe developmental defects including fused cotyledons, a short primary root, dwarf stature and defects in male fertility. We obtained evidence that PDK1 is responsible for AGC1 kinase PROTEIN KINASE ASSOCIATED WITH BRX (PAX) activation by phosphorylation during vascular development, and that the PDK1 phospholipid-binding Pleckstrin Homology domain is not required for this process. Our data indicate that PDK1 regulates polar auxin transport by activating AGC1 clade kinases, resulting in PIN phosphorylation.
3-磷酸肌醇依赖性蛋白激酶 1(PDK1)是真核生物中 AGC 激酶的保守主调控因子。pdk1 功能丧失会导致动物和酵母产生致命表型,但在拟南芥(Arabidopsis)中仅产生轻微的表型缺陷。拟南芥基因组包含两个 PDK1 编码基因,PDK1 和 PDK2。在这里,我们使用成簇规律间隔短回文重复(CRISPR)/CRISPR 相关蛋白 9(Cas9)生成真正的 PDK1 功能丧失等位基因,当与 pdk2 等位基因结合时,表现出严重的发育缺陷,包括融合的子叶、短主根、矮小身材和雄性育性缺陷。我们获得的证据表明,PDK1 通过在血管发育过程中磷酸化来负责 AGC1 激酶 PROTEIN KINASE ASSOCIATED WITH BRX(PAX)的激活,并且 PDK1 的磷脂结合 Pleckstrin Homology 结构域对于该过程不是必需的。我们的数据表明,PDK1 通过激活 AGC1 类激酶来调节极性生长素运输,导致 PIN 磷酸化。