Hill Joseph Lee, Josephs Cooper, Barnes William J, Anderson Charles T, Tien Ming
The Center for Lignocellulose Structure and Formation, The Pennsylvania State University, University Park, USA.
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, USA.
Plant Mol Biol. 2018 Feb;96(3):279-289. doi: 10.1007/s11103-017-0695-4. Epub 2018 Jan 31.
Our work focuses on understanding the lifetime and thus stability of the three main cellulose synthase (CESA) proteins involved in primary cell wall synthesis of Arabidopsis. It had long been thought that a major means of CESA regulation was via their rapid degradation. However, our studies here have uncovered that AtCESA proteins are not rapidly degraded. Rather, they persist for an extended time in the plant cell. Plant cellulose is synthesized by membrane-embedded cellulose synthase complexes (CSCs). The CSC is composed of cellulose synthases (CESAs), of which three distinct isozymes form the primary cell wall CSC and another set of three isozymes form the secondary cell wall CSC. We determined the stability over time of primary cell wall (PCW) CESAs in Arabidopsis thaliana seedlings, using immunoblotting after inhibiting protein synthesis with cycloheximide treatment. Our work reveals very slow turnover for the Arabidopsis PCW CESAs in vivo. Additionally, we show that the stability of all three CESAs within the PCW CSC is altered by mutations in individual CESAs, elevated temperature, and light conditions. Together, these results suggest that CESA proteins are very stable in vivo, but that their lifetimes can be modulated by intrinsic and environmental cues.
我们的工作重点是了解参与拟南芥初生细胞壁合成的三种主要纤维素合酶(CESA)蛋白的寿命,从而了解其稳定性。长期以来,人们一直认为CESA调节的主要方式是通过其快速降解。然而,我们在此的研究发现,拟南芥CESA蛋白不会快速降解。相反,它们在植物细胞中持续存在较长时间。植物纤维素由膜嵌入的纤维素合酶复合体(CSC)合成。CSC由纤维素合酶(CESA)组成,其中三种不同的同工型形成初生细胞壁CSC,另一组三种同工型形成次生细胞壁CSC。我们使用环己酰亚胺处理抑制蛋白质合成后的免疫印迹法,测定了拟南芥幼苗中初生细胞壁(PCW)CESA随时间的稳定性。我们的工作揭示了拟南芥PCW CESA在体内的周转非常缓慢。此外,我们表明,PCW CSC内所有三种CESA的稳定性会因单个CESA的突变、温度升高和光照条件而改变。总之,这些结果表明CESA蛋白在体内非常稳定,但其寿命可以受到内在和环境因素的调节。