Cytohesin dependent ARF to Rac signaling in HGF mediated motility

HGF 介导的运动中细胞粘附素依赖性 ARF 至 Rac 信号传导

• 批准号: 9132250
• 负责人: Lorraine C Santy
• 金额: $ 29.05万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-20 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9132250
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Affect; Area; Binding; Cell Shape; Cell Survival; Cells; Cessation of life; Complication; Cytosol; Data; Development; Epithelial; Epithelial Cells; Event; Fibrosis; Goals; Growth; Guanosine Triphosphate Phosphohydrolases; HGF gene; Hospitals; Immigration; Injury; Kidney; Knowledge; Link; Lipids; Mediating; Membrane; Modeling; Modification; Molecular; Molecular Conformation; Monomeric GTP-Binding Proteins; Movement; Mutagenesis; Neoplasm Metastasis; Normal Cell; Pathway interactions; Patients; Phosphatidylinositols; Phosphorylation; Process; Production; Publishing; Recovery; Recruitment Activity; Renal function; Reperfusion Injury; Research; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Small Interfering RNA; Staging; Surface; Testing; Work; Wound Healing; base; cell motility; fighting; kidney repair; knock-down; migration; mutant; prevent; protein protein interaction; renal ischemia; repaired; research study; response; scaffold; therapeutic target; therapy design; therapy development; tool development; tumor

DESCRIPTION (provided by applicant): Acute kidney injury is a common hospital complication that leads death or long-term kidney damage if recovery is not complete. Hepatocyte growth factor (HGF) promotes the recovery from kidney damage at multiple steps. While many of the proximal signals activated by HGF and the downstream targets that remodel cell shape in response to HGF are known, the details of the circuits that link them together are unclear. This limits the possibility of selectively modulating HGF signaling without impacting the actions of other growth factors. Work in the applicant's lab has recently demonstrated that protein- protein interactions are critical for building a pro-migratory circuit that links activatin of Arf6 by cytohesin-2 to the downstream activation of Rac by Dock180. Preliminary data suggest that this signaling circuit is required for HGF-stimulated Rac activation and migration. The overall objective of this application is to determine the molecular details of this cytohesin-dependent Arf-to-Rac signaling module and to test the degree to which it mediates HGF-induced motility and the recovery from acute kidney injury. This is the next step toward the long-term goal of understanding the initiation of motility in epithelial cells by small GTPase cascades. This application will test a hypothesized model for the cytohesin-dependent Arf-to-Rac signaling module. This module will be tested by pursuing 4 specific aims: 1) Determine how cytohesin-2 activity is controlled; 2) Determine the mechanism of interaction of cytohesin-2 and Dock180 and test the degree to which this interaction mediates HGF dependent Rac activation; 3) Determine the role of active Arf6 in cytohesin-dependent Rac activation; 4) Determine the extent to which cytohesin-dependent Rac activation regulates HGF-stimulated wound healing and recovery of the kidney from ischemia reperfusion injury. Aim 1 will test the hypothesis that cytohesin-2 intramolecular interactions prevent its interaction with membranes until these interactions are disrupted by phosphorylation. Aim 2 will use mutagenesis to identify the regions necessary for the interaction of the cytohesin-binding scaffold GRASP with Dock180. The importance of this interaction for HGF- induced Rac activation will be tested using knockdown of GRASP and expression of GRASP mutants. Aim 3 will test the hypothesis that Arf6-dependent lipid modifications promote activation of Rac by Dock180. Aim 4 will use siRNA, mutant and pharmacologic modulation of cytohesin-dependent Arf-to- Rac signaling to test its involvement in HGF-stimulated wound healing and recovery of the kidney from ischemia reperfusion injury. A proven model of this pro-migratory circuit will allow the design of interventions to modulate this circuit without impacting other Arf6 or Rac-dependent functions. Such treatments could promote or inhibit HGF-dependent migration without disrupting the actions of a broad range of growth factors.

描述（由申请人提供）：急性肾损伤是一种常见的医院并发症，如果恢复不完全，可导致死亡或长期肾损害。肝细胞生长因子（HGF）通过多个步骤促进肾损伤的恢复。虽然已知许多由HGF激活的近端信号和响应HGF重塑细胞形状的下游目标，但将它们连接在一起的电路的细节尚不清楚。这限制了在不影响其他生长因子作用的情况下选择性调节HGF信号的可能性。申请人实验室最近的工作表明，蛋白质-蛋白质相互作用对于构建一个促进迁移的回路至关重要，该回路将细胞素-2对Arf6的激活与Dock180对Rac的下游激活联系起来。初步数据表明，这种信号通路是hgf刺激的Rac激活和迁移所必需的。本应用程序的总体目标是确定这种依赖于细胞染色素的Arf-to-Rac信号传导模块的分子细节，并测试其介导hgf诱导的运动和急性肾损伤恢复的程度。这是下一步的长期目标，了解启动运动在上皮细胞的小GTPase级联。

项目成果

The cytohesin coiled-coil domain interacts with threonine 276 to control membrane association.

• DOI: 10.1371/journal.pone.0082084
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Hiester KG;Santy LC
• 通讯作者: Santy LC

The scaffolding protein GRASP/Tamalin directly binds to Dock180 as well as to cytohesins facilitating GTPase crosstalk in epithelial cell migration.

• DOI: 10.1186/1471-2121-14-9
• 发表时间: 2013-02-26
• 期刊: BMC cell biology
• 作者: Attar MA;Santy LC
• 通讯作者: Santy LC

A traction force threshold signifies metastatic phenotypic change in multicellular epithelia.

• DOI: 10.1039/c9sm00733d
• 发表时间: 2019-09
• 期刊: Soft matter
• 影响因子: 3.4
• 作者: Yao Zhang;Xuechen Shi;Tiankai Zhao;Changjin Huang;Qiong Wei;Xin Tang;Lorraine C. Santy;M. Taher A. Saif;Sulin Zhang

ARF-GEF cytohesin-2/ARNO regulates R-Ras and α5-integrin recycling through an EHD1-positive compartment.

• DOI: 10.1091/mbc.e15-05-0278
• 发表时间: 2015-11-15
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Salem JC;Reviriego-Mendoza MM;Santy LC
• 通讯作者: Santy LC

The cytohesin guanosine exchange factors (GEFs) are required to promote HGF-mediated renal recovery after acute kidney injury (AKI) in mice.

• DOI: 10.14814/phy2.12442
• 发表时间: 2015-06
• 期刊: Physiological reports
• 影响因子: 2.5
• 作者: Reviriego-Mendoza MM;Santy LC
• 通讯作者: Santy LC