Activation of actin-depolymerizing factor by CDPK16-mediated phosphorylation promotes actin turnover in Arabidopsis pollen tubes.
Activation of actin-depolymerizing factor by CDPK16-mediated phosphorylation promotes actin turnover in Arabidopsis pollen tubes.
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DOI：
10.1371/journal.pbio.3002073
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发表时间：
2023-04
期刊：
影响因子：
9.8
通讯作者：
Huang, Shanjin
中科院分区：
文献类型：
--
作者：
Wang, Qiannan;Xu, Yanan;Zhao, Shuangshuang;Jiang, Yuxiang;Yi, Ran;Guo, Yan;Huang, Shanjin
作为肌动蛋白动态变化的刺激响应调节因子，肌动蛋白解聚因子（ADF）/丝切蛋白受到严格调控。众所周知，激酶介导的磷酸化会使ADF/丝切蛋白失活。然而，在此我们发现，拟南芥ADF7的活性会因CDPK16介导的磷酸化而增强。我们发现，CDPK16在体外和体内均能与ADF7相互作用，并且它能以钙依赖的方式在体外增强ADF7介导的肌动蛋白解聚和切断作用。相应地，cdpk16突变体花粉中肌动蛋白的周转速率降低，且cdpk16突变体花粉管顶端的肌动蛋白丝数量显著增加。CDPK16在体外和体内均可使ADF7的丝氨酸128位点发生磷酸化，与ADF7相比，模拟磷酸化突变体ADF7S128D具有更强的肌动蛋白解聚活性。引人注目的是，我们发现ADF7丝氨酸128位点无法发生磷酸化会损害其在体内促进肌动蛋白周转的功能，这表明这种磷酸化调控机制具有重要的生物学意义。因此，我们揭示了CDPK16介导的磷酸化作用会上调ADF7，从而促进花粉中的肌动蛋白周转。
作为肌动蛋白动态变化的刺激响应调节因子，ADF/丝切蛋白受到严格调控。本研究表明，拟南芥ADF7的肌动蛋白解聚和切断活性因CDPK16介导的丝氨酸128位点磷酸化而增强，进而促进花粉中的肌动蛋白周转。
As the stimulus-responsive mediator of actin dynamics, actin-depolymerizing factor (ADF)/cofilin is subject to tight regulation. It is well known that kinase-mediated phosphorylation inactivates ADF/cofilin. Here, however, we found that the activity of Arabidopsis ADF7 is enhanced by CDPK16-mediated phosphorylation. We found that CDPK16 interacts with ADF7 both in vitro and in vivo, and it enhances ADF7-mediated actin depolymerization and severing in vitro in a calcium-dependent manner. Accordingly, the rate of actin turnover is reduced in cdpk16 pollen and the amount of actin filaments increases significantly at the tip of cdpk16 pollen tubes. CDPK16 phosphorylates ADF7 at Serine128 both in vitro and in vivo, and the phospho-mimetic mutant ADF7S128D has enhanced actin-depolymerizing activity compared to ADF7. Strikingly, we found that failure in the phosphorylation of ADF7 at Ser128 impairs its function in promoting actin turnover in vivo, which suggests that this phospho-regulation mechanism is biologically significant. Thus, we reveal that CDPK16-mediated phosphorylation up-regulates ADF7 to promote actin turnover in pollen. As the stimulus-responsive mediator of actin dynamics, ADF/cofilin is tightly regulated. This study shows that the actin depolymerization and severing activity of Arabidopsis ADF7 is enhanced by CDPK16-mediated phosphorylation at Ser128 to promote actin turnover in pollen.
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