Wiesler Bodo, Wang Qi-Yan, Nick Peter
Institut für Biologie II/Botanik, Schänzlestrasse 1, Universität Freiburg, D-79104 Freiburg, Germany.
Plant J. 2002 Dec;32(6):1023-32. doi: 10.1046/j.1365-313x.2002.01489.x.
Auxin controls the orientation of cortical microtubules in maize coleoptile segments. We used tyrosinylated alpha-tubulin as a marker to assess auxin-dependent changes in microtubule turnover. Auxin-induced tyrosinylated alpha-tubulin, correlated with an elevated sensitivity of growth to antimicrotubular compounds such as ethyl-N-phenylcarbamate (EPC). We determined the affinity of alpha-tubulin to EPC and found that it was dramatically increased when the tubulin was de-tyrosinylated. By proteolytic cleavage of the carboxy terminal tyrosine, such an increased affinity could be induced in vitro. Thus, the auxin-induced sensitivity of growth to EPC is not caused by an increased affinity for this inhibitor, but caused by a reduced microtubule turnover. Double visualization assays revealed that the transverse microtubules induced by auxin consist predominantly of tyrosinylated alpha-tubulin, whereas the longitudinal microtubules induced by auxin depletion contain de-tyrosinylated alpha-tubulin. The results are discussed in terms of direction-dependent differences in the lifetime of microtubules.
生长素控制玉米胚芽鞘切段中皮层微管的取向。我们使用酪氨酸化α-微管蛋白作为标记物来评估生长素依赖性的微管周转变化。生长素诱导的酪氨酸化α-微管蛋白,与生长对诸如乙基-N-苯基氨基甲酸酯(EPC)等抗微管化合物的敏感性升高相关。我们测定了α-微管蛋白对EPC的亲和力,发现当微管蛋白去酪氨酸化时,其亲和力显著增加。通过羧基末端酪氨酸的蛋白水解切割,这种增加的亲和力可以在体外诱导产生。因此,生长素诱导的生长对EPC的敏感性不是由对该抑制剂的亲和力增加引起的,而是由微管周转减少引起的。双重可视化分析表明,生长素诱导的横向微管主要由酪氨酸化α-微管蛋白组成,而生长素耗尽诱导的纵向微管含有去酪氨酸化α-微管蛋白。根据微管寿命的方向依赖性差异对结果进行了讨论。