Regulation of GTPase function by autophosphorylation.
Regulation of GTPase function by autophosphorylation.
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DOI：
10.1016/j.molcel.2022.02.011
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发表时间：
2022-03-03
期刊：
影响因子：
16
通讯作者：
Haigis KM
中科院分区：
文献类型：
--
作者：
Johnson CW;Seo HS;Terrell EM;Yang MH;KleinJan F;Gebregiworgis T;Gasmi-Seabrook GMC;Geffken EA;Lakhani J;Song K;Bashyal P;Popow O;Paulo JA;Liu A;Mattos C;Marshall CB;Ikura M;Morrison DK;Dhe-Paganon S;Haigis KM
RAS超家族的一个统一特征是保守的GTdR循环，这些蛋白质通过该循环在活性和非活性状态之间转变。我们证明，一些GTP酶的自磷酸化是一种内在的调节机制，减少核苷酸水解和增强核苷酸交换，改变开/关开关，形成其信号功能的基础。使用X-射线晶体学，核磁共振光谱，结合分析，和分子动力学上的自磷酸化突变体的H-RAS和K-RAS，我们表明，磷酸转移从GTP需要动态运动的开关II区和自磷酸化促进核苷酸交换，通过打开活性位点和提取稳定Mg 2+。最后，我们证明了自磷酸化的K-RAS表现出改变的效应相互作用，包括在哺乳动物细胞中RAF蛋白的亲和力降低。因此，自磷酸化导致改变的活性位点动力学和效应物相互作用性质，产生一个在功能上不同于其非磷酸化对应物的GTP酶库。约翰逊等人鉴定了一组通过保守的活性位点取代进行自磷酸化的GTP酶。使用RASA 59 T作为原型自磷酸化GTP酶，他们表明自磷酸化是一种稳定的翻译后修饰，可抑制GTP水解并增强核苷酸交换。尽管促进细胞转化，但自磷酸化抑制K-RAS效应物相互作用。
A unifying feature of the RAS superfamily is a conserved GTPase cycle by which these proteins transition between active and inactive states. We demonstrate that autophosphorylation of some GTPases is an intrinsic regulatory mechanism that reduces nucleotide hydrolysis and enhances nucleotide exchange, altering the on/off switch that forms the basis for their signaling functions. Using X-ray crystallography, nuclear magnetic resonance spectroscopy, binding assays, and molecular dynamics on autophosphorylated mutants of H-RAS and K-RAS, we show that phosphoryl transfer from GTP requires dynamic movement of the switch II region and that autophosphorylation promotes nucleotide exchange by opening the active site and extracting the stabilizing Mg2+. Finally, we demonstrate that autophosphorylated K-RAS exhibits altered effector interactions, including a reduced affinity for RAF proteins in mammalian cells. Thus, autophosphorylation leads to altered active site dynamics and effector interaction properties, creating a pool of GTPases that are functionally distinct from their non-phosphorylated counterparts. Johnson et al. identify a group of GTPases that undergo autophosphorylation via a conserved active site substitution. Using RASA59T as a prototypical autophosphorylating GTPase, they show that autophosphorylation is a stable post-translational modification that inhibits GTP hydrolysis and enhances nucleotide exchange. Despite promoting cell transformation, autophosphorylation inhibits K-RAS effector interactions.
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影响因子：
14.9
作者：
NCBI Resource Coordinators
通讯作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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DOI：
10.1107/s0907444909052925
发表时间：
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期刊：
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影响因子：
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作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH