LIM domain kinases: regulation and substrate recognition

LIM 结构域激酶：调节和底物识别

• 批准号: 10798525
• 负责人: Titus Jonathon Boggon
• 金额: $ 9.36万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798525
• 关键词: Actins; Adaptor Signaling Protein; Architecture; Area; Binding; Binding Proteins; Biochemical; Biophysics; CDC42 gene; Cells; Cellular Morphology; Complex; Coupled; Cues; Cytoskeleton; Data; Development; Enzymes; Eukaryotic Cell; Family; Guanosine Triphosphate Phosphohydrolases; Homeostasis; LIM Domain; LIM Domain Kinase 1; LIMK1 gene; Laboratories; Mediating; Microfilaments; Modeling; Molecular; Movement; Myotonic Dystrophy; Phosphorylation; Phosphotransferases; Process; Protein Kinase; Proteins; Regulation; Signal Pathway; Signal Transduction; Specificity; Surface; X-Ray Crystallography; actin depolymerizing factor; cell growth regulation; cofilin; extracellular; molecular dynamics; p21 activated kinase; recruit; rho GTP-Binding Proteins; upstream kinase

Multi-PI: Titus J. Boggon and Benjamin E. Turk LIM Domain Kinases: Regulation and Substrate Recognition ABSTRACT Eukaryotic cells interpret extracellular and intrinsic cues to effect remodeling of the actin cytoskeleton, a process critical for controlling cell morphology, movement, and invasiveness. Tight control of signaling pathways impinging on the cytoskeleton is therefore essential to normal development and homeostasis. The RHO family GTPases RHO, RAC and CDC42 each directly activate kinases (RHO kinases, PAKs, and MRCKs) in a spatially restricted manner that in turn directly phosphorylate and activate the LIM domain kinases (LIMK1 and LIMK2). These kinase signaling cascades ultimately converge on phosphorylation of the cofilin/ADF (actin- depolymerizing factor) group of proteins, key molecules that mediate remodeling of actin filaments. Over the previous two periods we have leveraged the highly complementary expertise of our two laboratories to provide significant advances in two main areas: understanding the specificity and regulation of p21-activated kinases (PAKs) and revealing the basis for selective targeting of cofilin by LIMKs. We will now target our efforts toward answering outstanding questions that remain regarding regulation and function of LIMKs. Our preliminary data suggest that an intramolecular interaction between a LIM-PDZ module and the kinase domain, potentially involving evolutionarily conserved binding surfaces, is responsible for suppressing LIMK catalytic activity. Combining biophysical, biochemical, and cell-based approaches, we will address the hypothesis that disruption of this interaction results in activation of the LIM kinases, and we will reveal the structural basis for LIMK autoregulation. We will further investigate recent evidence that LIMKs can phosphorylate both Ser and Tyr residues by X-ray crystallography of LIMK-substrate complexes and molecular dynamics simulations. In this way LIMKs will serve as a general model for understanding substrate recognition by the various “dual specificity” kinase families. Finally, we will investigate the myotonic dystrophy related CDC42-binding protein kinases (MRCKs), a major group of LIMK activating kinases downstream of the GTPase CDC42, about which little is currently known. We will use structural, biophysical and biochemical approaches to define the basic architecture of MRCKβ studies and to probe how its activation is coupled to interactions with LIMKs through substrate adaptor proteins. Overall, our studies will provide a substantial advance in our molecular level understanding of signaling pathways downstream of the RHO family GTPases that impinge on regulation of the actin cytoskeleton.

多元派：泰特斯·J·博根和本杰明·E·特克 LIM结构域蛋白激酶：调控和底物识别 摘要 真核细胞解释细胞外和内在的信号来影响肌动蛋白细胞骨架的重塑，这是一个过程 对控制细胞形态、运动和侵袭性至关重要。严格控制信号通路 因此，对细胞骨架的冲击对正常发育和动态平衡是必不可少的。罗氏家族 GTP酶Rho、Rac和CDC42中的每一个都直接激活空间上 LIM结构域激酶(LIMK1和LIMK2)被限制的方式直接磷酸化并激活。 这些信号通路最终汇聚在cofilin/ADF(肌动蛋白-肌动蛋白-肌动蛋白)的磷酸化过程中。 解聚因子)一组蛋白质，调节肌动蛋白细丝重塑的关键分子。超过了 在前两个阶段，我们利用我们两个实验室高度互补的专业知识来提供 两个主要领域的重大进展：了解p21激活的激酶的特异性和调节 (PAKS)，并揭示了LIMK选择性靶向Cofilin的基础。我们现在将把我们的努力集中在 回答关于LIMK的监管和功能的悬而未决的问题。我们的初步数据 提示LIM-PDZ模块和激酶结构域之间的分子内相互作用，可能 涉及进化上保守的结合表面，是抑制LIMK催化活性的原因。 结合生物物理、生化和基于细胞的方法，我们将解决破坏的假设 这种相互作用导致LIM激酶的激活，我们将揭示LIMK的结构基础 自动调节。我们将进一步调查最近的证据，即LIMK可以同时磷酸化丝氨酸和酪氨酸 LIMK底物络合物的X-射线结晶学和分子动力学模拟。以这种方式 LIMKs将作为理解底物识别的一般模型，通过各种“双重专一性” 激活酶家族。最后，我们将研究与cdc42结合蛋白激酶相关的强直性肌营养不良。 (MRCKs)，是GTPase CDc42下游的一组主要LIMK激活蛋白激酶，关于它的研究很少 目前已知的。我们将使用结构、生物物理和生化方法来定义基本架构 研究并探索其激活如何通过底物适配器与LIMK相互作用 蛋白质。总体而言，我们的研究将为我们在分子水平上理解信号提供实质性的进步 Rho家族GTP酶下游影响肌动蛋白细胞骨架调控的途径。

项目成果

TNIK Is a Therapeutic Target in Lung Squamous Cell Carcinoma and Regulates FAK Activation through Merlin.

• DOI: 10.1158/2159-8290.cd-20-0797
• 发表时间: 2021-06
• 期刊: Cancer discovery
• 影响因子: 28.2
• 作者: Torres-Ayuso P;An E;Nyswaner KM;Bensen RC;Ritt DA;Specht SI;Das S;Andresson T;Cachau RE;Liang RJ;Ries AL;Robinson CM;Difilippantonio S;Gouker B;Bassel L;Karim BO;Miller CJ;Turk BE;Morrison DK;Brognard J
• 通讯作者: Brognard J

PseudoGTPase domains in p190RhoGAP proteins: a mini-review.

p190RhoGAP 蛋白中的伪 GTP 酶结构域：小型综述。

• DOI: 10.1042/bst20180481
• 发表时间: 2018
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Stiegler,AmyL;Boggon,TitusJ
• 通讯作者: Boggon,TitusJ

Signaling pathways and the cerebral cavernous malformations proteins: lessons from structural biology.

• DOI: 10.1007/s00018-013-1532-9
• 发表时间: 2014-05
• 期刊: CELLULAR AND MOLECULAR LIFE SCIENCES
• 影响因子: 8
• 作者: Fisher, Oriana S.;Boggon, Titus J.
• 通讯作者: Boggon, Titus J.

Homing in: Mechanisms of Substrate Targeting by Protein Kinases.

• DOI: 10.1016/j.tibs.2018.02.009
• 发表时间: 2018-05
• 期刊: Trends in biochemical sciences
• 影响因子: 13.8
• 作者: Miller CJ;Turk BE
• 通讯作者: Turk BE

Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail.

独特的功能限制驱动丝切蛋白 N 末端调控尾部的保护。

• DOI: 10.1101/2023.06.30.547189
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Sexton,JoelA;Potchernikov,Tony;Bibeau,JeffreyP;Casanova-Sepúlveda,Gabriela;Cao,Wenxiang;Lou,HuaJane;Boggon,TitusJ;DeLaCruz,EnriqueM;Turk,BenjaminE
• 通讯作者: Turk,BenjaminE