Multiple phosphorylation sites on the RegA phosphodiesterase regulate Dictyostelium development.
Multiple phosphorylation sites on the RegA phosphodiesterase regulate Dictyostelium development.
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RegA 磷酸二酯酶上的多个磷酸化位点调节盘基网柄菌的发育。
DOI：
10.1016/j.cellsig.2019.02.005
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发表时间：
2019
影响因子：
4.8
通讯作者：
Hadwiger,JeffreyA
中科院分区：
文献类型：
--
作者：
Kuburich,NickA;Adhikari,Nirakar;Hadwiger,JeffreyA
在网骨藻中，细胞内cAMP特异性磷酸二酯酶RegA是cAMP依赖性蛋白激酶（PKA）的负调节剂，PKA是发育形态发生和孢子形成时间的关键决定因素。为了评估蛋白激酶在调节RegA功能中的作用，本研究确定了RegA上的磷酸化位点，并通过分析磷酸模拟和磷酸消融突变来表征这些修饰的作用。影响RegA残基T676（非典型MAP激酶Erk2的假定靶点）的突变改变了嵌合体的发育速率并影响了细胞分布，这表明该残基的磷酸化降低了RegA功能并调节了多细胞发育过程中的细胞定位。影响RegA残基S142的突变也影响发育形态发生的速率，但与T676的变化相反，表明S142残基的磷酸化增加了RegA功能。影响残基S413残基的突变改变了聚集体大小并延迟了发育进程，这表明PKA以负反馈机制来增加RegA功能。这些结果表明，RegA上不同残基的磷酸化可以导致RegA功能的增加或减少，从而反过来调节发育过程，如聚集体形成，细胞分布和发育形态发生的动力学。
In Dictyostelium, the intracellular cAMP-specific phosphodiesterase RegA is a negative regulator of cAMP-dependent protein kinase (PKA), a key determinant in the timing of developmental morphogenesis and spore formation. To assess the role of protein kinases in the regulation of RegA function, this study identified phosphorylation sites on RegA and characterized the role of these modifications through the analysis of phospho-mimetic and phospho-ablative mutations. Mutations affecting residue T676 of RegA, a presumed target of the atypical MAP kinase Erk2, altered the rate of development and impacted cell distribution in chimeric organisms suggesting that phosphorylation of this residue reduces RegA function and regulates cell localization during multicellular development. Mutations affecting the residue S142 of RegA also impacted the rate developmental morphogenesis but in a manner opposite of changes at T676 suggesting the phosphorylation of the S142 residue increases RegA function. Mutations affecting residue S413 residue altered aggregate sizes and delayed developmental progression suggesting that PKA operates in a negative feedback mechanism to increase RegA function. These results suggest that the phosphorylation of different residues on RegA can lead to increased or decreased RegA function and therefore in turn regulate developmental processes such as aggregate formation, cell distribution, and the kinetics of developmental morphogenesis.