Department of Botany, Graduate School of Science, Kyoto University, Kyoto, Japan.
Plant Cell Physiol. 2011 Sep;52(9):1628-40. doi: 10.1093/pcp/pcr093. Epub 2011 Jul 22.
The 26S proteasome plays fundamental roles in the degradation of short-lived regulatory proteins, thereby controlling diverse cellular processes. In Arabidopsis, the essential RPT2 subunit is encoded by two highly homologous genes: RPT2a and RPT2b. Currently, only RPT2a has been reported to regulate various developmental processes, including the maintenance of the root apical meristem (RAM), although the roles of RPT2a in the RAM are still obscure. Here, we analyzed the cell type-specific requirement for RPT2a. When RPT2a was expressed locally in the rpt2a mutant, pleiotropic defects in the RAM, such as cell death and distorted cellular organization, were rescued differently, suggesting that RPT2a regulates various specific activities, which converge to maintain the RAM. On the other hand, the homologous RPT2b was also expressed in meristems, and the expression of RPT2b protein under the control of the RPT2a promoter complemented the rpt2a RAM defects, although the rpt2b mutant showed no obvious defect in all developmental aspects we examined. These results show that RPT2b might work in the RAM, but is dispensable for RAM maintenance in the presence of RPT2a. In contrast, the rpt2a rpt2b double mutant was lethal in male and female gametophytes, suggesting that RPT2a and RPT2b are redundantly required for gametogenesis. Furthermore, we showed that similar meristematic and gametophytic defects were caused by mutations in other subunit genes, RPT5a and RPT5b, suggesting that proper activity of the proteasome, not an RPT2-specific function, is required. Taken together, our results suggest that RPT2a and RPT2b contribute differently to the proteasome activity required for each developmental context.
26S 蛋白酶体在降解短寿命调节蛋白中起着至关重要的作用,从而控制着多种细胞过程。在拟南芥中,必需的 RPT2 亚基由两个高度同源的基因编码:RPT2a 和 RPT2b。目前,只有 RPT2a 被报道调节各种发育过程,包括维持根分生组织(RAM),尽管 RPT2a 在 RAM 中的作用仍然不清楚。在这里,我们分析了 RPT2a 在细胞类型特异性中的需求。当 RPT2a 在 rpt2a 突变体中局部表达时,RAM 中的多效缺陷,如细胞死亡和细胞组织变形,得到了不同程度的挽救,这表明 RPT2a 调节着各种特定的活动,这些活动汇聚在一起维持 RAM。另一方面,同源的 RPT2b 也在分生组织中表达,在 RPT2a 启动子控制下表达的 RPT2b 蛋白可以弥补 rpt2a 的 RAM 缺陷,尽管在我们检查的所有发育方面,rpt2b 突变体都没有明显的缺陷。这些结果表明,RPT2b 可能在 RAM 中发挥作用,但在存在 RPT2a 的情况下,对于 RAM 的维持是可有可无的。相比之下,rpt2a rpt2b 双突变体在雄性和雌性配子体中是致命的,这表明 RPT2a 和 RPT2b 在配子发生中是冗余的。此外,我们还表明,其他亚基基因 RPT5a 和 RPT5b 的突变也会导致类似的分生组织和配子体缺陷,这表明蛋白酶体的适当活性,而不是 RPT2 的特定功能,是必需的。综上所述,我们的结果表明,RPT2a 和 RPT2b 在每个发育背景下对蛋白酶体活性的贡献不同。