LIM domain kinases: regulation and substrate recognition

LIM 结构域激酶：调节和底物识别

基本信息

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
项目类别：
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

项目摘要

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.

Multi-Pi：Titus J. Boggon和Benjamin E. Turk LIM域激酶：调节和底物识别抽象的真核细胞解释细胞外和内在提示以实现肌动蛋白细胞骨架的重塑，这是一种过程控制细胞形态，运动和侵入性至关重要。信号通路的严格控制因此，撞击细胞骨架对于正常发育和稳态至关重要。 Rho家族 GTPASE RHO，RAC和CDC42各自直接激活激酶（Rho激酶，PAKS和MRCKS）限制了直接磷酸化并激活LIM结构域激酶（Limk1和Limk2）的限制方式。这些激酶信号传导级联对于Cofilin/adf的磷酸化最终（肌动蛋白 - 解聚因子）蛋白质组，介导肌动蛋白丝的重塑的关键分子。在前两个时期，我们利用了两个实验室的高度完善专业知识来提供在两个主要领域的重大进展：了解P21激活激酶的特异性和调节（paks）并揭示了通过四肢选择性靶向cofilin的基础。现在，我们将把我们的努力定向回答有关肢体的调节和功能仍然存在的杰出问题。我们的初步数据表明LIM-PDZ模块与激酶结构域之间的分子内相互作用，可能有可能涉及进化保守的结合表面，负责抑制limk催化活性。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，我们将揭示肢体的结构基础自动调节。我们将进一步调查最近的证据表明四肢可以磷酸化Ser和Tyr 通过X射线晶体学和分子动力学模拟的X射线晶体学的残基。这样 LIMK将成为通过各种“双重特异性”理解底物识别的一般模型激酶家庭。最后，我们将研究与肌营养不良症相关的CDC42结合蛋白激酶（MRCKS），一组主要的肢体激活GTPase Cdc42下游激活激酶，几乎没有什么是目前已知。我们将使用结构，生物物理和生化方法来定义基本体系结构 MRCKβ研究并探测其激活如何通过底物适配器耦合与四肢的相互作用蛋白质。总体而言，我们的研究将在我们对信号传导的分子水平理解中提供重大进步 Rho家族GTPases下游的途径会影响肌动蛋白细胞骨架的调节。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
影响因子：
0
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