Department of Crop and Soil Sciences, Washington State University, Pullman, Washington, USA.
Molecular Plant Sciences, Washington State University, Pullman, Washington, USA.
Plant Physiol. 2022 Nov 28;190(4):2651-2670. doi: 10.1093/plphys/kiac406.
The plant Ubiquitin Regulatory X (UBX) domain-containing protein 1 (PUX1) functions as a negative regulator of gibberellin (GA) signaling. GAs are plant hormones that stimulate seed germination, the transition to flowering, and cell elongation and division. Loss of Arabidopsis (Arabidopsis thaliana) PUX1 resulted in a "GA-overdose" phenotype including early flowering, increased stem and root elongation, and partial resistance to the GA-biosynthesis inhibitor paclobutrazol during seed germination and root elongation. Furthermore, GA application failed to stimulate further stem elongation or flowering onset suggesting that elongation and flowering response to GA had reached its maximum. GA hormone partially repressed PUX1 protein accumulation, and PUX1 showed a GA-independent interaction with the GA receptor GA-INSENSITIVE DWARF-1 (GID1). This suggests that PUX1 is GA regulated and/or regulates elements of the GA signaling pathway. Consistent with PUX1 function as a negative regulator of GA signaling, the pux1 mutant caused increased GID1 expression and decreased accumulation of the DELLA REPRESSOR OF GA1-3, RGA. PUX1 is a negative regulator of the hexameric AAA+ ATPase CDC48, a protein that functions in diverse cellular processes including unfolding proteins in preparation for proteasomal degradation, cell division, and expansion. PUX1 binding to GID1 required the UBX domain, a binding motif necessary for CDC48 interaction. Moreover, PUX1 overexpression in cell culture not only stimulated the disassembly of CDC48 hexamer but also resulted in co-fractionation of GID1, PUX1, and CDC48 subunits in velocity sedimentation assays. Based on our results, we propose that PUX1 and CDC48 are additional factors that need to be incorporated into our understanding of GA signaling.
植物泛素调节 X(UBX)结构域蛋白 1(PUX1)作为赤霉素(GA)信号的负调节剂发挥作用。GA 是一种植物激素,可刺激种子萌发、向花的转变以及细胞伸长和分裂。拟南芥(Arabidopsis thaliana)PUX1 的缺失导致了“GA 过量”表型,包括早花、茎和根伸长增加以及在种子萌发和根伸长过程中对 GA 生物合成抑制剂多效唑的部分抗性。此外,GA 处理未能刺激茎进一步伸长或开花启动,表明对 GA 的伸长和开花反应已达到最大。GA 激素部分抑制 PUX1 蛋白积累,并且 PUX1 与 GA 受体 GA-INSENSITIVE DWARF-1(GID1)表现出 GA 独立的相互作用。这表明 PUX1 受 GA 调节和/或调节 GA 信号通路的元件。与 PUX1 作为 GA 信号的负调节剂的功能一致,pux1 突变体导致 GID1 表达增加和 DELLA 抑制物 GA1-3、RGA 的积累减少。PUX1 是六聚体 AAA+ATP 酶 CDC48 的负调节剂,CDC48 在包括蛋白质变性以准备蛋白酶体降解、细胞分裂和扩张在内的多种细胞过程中发挥作用。PUX1 与 GID1 的结合需要 UBX 结构域,这是与 CDC48 相互作用的必需结合基序。此外,细胞培养中的 PUX1 过表达不仅刺激了 CDC48 六聚体的解体,而且还导致在速度沉降测定中 GID1、PUX1 和 CDC48 亚基的共分馏。基于我们的结果,我们提出 PUX1 和 CDC48 是需要纳入我们对 GA 信号理解的另外两个因素。