Phosphorylation of the WH2 domain in yeast Las17/WASP regulates G-actin binding and protein function during endocytosis.
Phosphorylation of the WH2 domain in yeast Las17/WASP regulates G-actin binding and protein function during endocytosis.
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DOI：
10.1038/s41598-021-88826-z
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发表时间：
2021-05-06
影响因子：
4.6
通讯作者：
Ayscough KR
中科院分区：
文献类型：
--
作者：
Tyler JJ;Smaczynska-de Rooij II;Abugharsa L;Palmer JS;Hancock LP;Allwood EG;Ayscough KR
肌动蛋白成核是肌动蛋白聚合过程中关键的限速步骤，对该过程的严格调节对于确保肌动蛋白丝仅在特定时间和细胞的特定区域形成至关重要。 WH2 结构域是在许多不同的肌动蛋白结合蛋白中发现的短序列基序，包括调节肌动蛋白成核复合物 Arp2/3 的 WASP 家族蛋白。在这项研究中，我们揭示了酵母 WASP 同源物 Las17 的 WH2 结构域内的磷酸化位点丝氨酸 554。磷酸化和磷酸模拟突变都会降低肌动蛋白单体的结合亲和力，而模拟非磷酸化状态的丙氨酸突变会增加肌动蛋白的结合亲和力。分析了这些突变对体内 Las17 依赖的内吞作用过程的影响，并促使我们提出，Las17 磷酸化状态的转换可能允许从位点组装到最终分裂阶段经历不同的内吞作用阶段。虽然这项研究的重点是酵母中唯一的 WASP 家族蛋白 Las17，但我们的结果对于我们理解这一保守基序中的关键残基如何支持与 WH2 结构域相关的许多不同肌动蛋白调节作用具有广泛的意义。
Actin nucleation is the key rate limiting step in the process of actin polymerization, and tight regulation of this process is critical to ensure actin filaments form only at specific times and at defined regions of the cell. WH2 domains are short sequence motifs found in many different actin binding proteins including WASP family proteins which regulate the actin nucleating complex Arp2/3. In this study we reveal a phosphorylation site, Serine 554, within the WH2 domain of the yeast WASP homologue Las17. Both phosphorylation and a phospho-mimetic mutation reduce actin monomer binding affinity while an alanine mutation, generated to mimic the non-phosphorylated state, increases actin binding affinity. The effect of these mutations on the Las17-dependent process of endocytosis in vivo was analysed and leads us to propose that switching of Las17 phosphorylation states may allow progression through distinct phases of endocytosis from site assembly through to the final scission stage. While the study is focused on Las17, the sole WASP family protein in yeast, our results have broad implications for our understanding of how a key residue in this conserved motif can underpin the many different actin regulatory roles with which WH2 domains have been associated.
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DOI：
10.1083/jcb.144.6.1203
发表时间：
1999-03-22
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cope MJ;Yang S;Shang C;Drubin DG
通讯作者：
Drubin DG
影响因子：
9.2
作者：
Palmer, Sarah E.;Smaczynska-de Rooij, Iwona I.;Marklew, Christopher J.;Allwood, Ellen G.;Mishra, Ritu;Johnson, Simeon;Goldberg, Martin W.;Ayscough, Kathryn R.
通讯作者：
Ayscough, Kathryn R.
影响因子：
64.5
作者：
Kaksonen, M;Toret, CP;Drubin, DG
通讯作者：
Drubin, DG
影响因子：
64.5
作者：
Co, Carl;Wong, Derek T.;Taunton, Jack
通讯作者：
Taunton, Jack
影响因子：
16
作者：
Cory, GOC;Cramer, R;Ridley, AJ
通讯作者：
Ridley, AJ