Cortactin function in lamellipodial protrusion

Cortactin 在板状足突出中的功能

• 批准号: 8118809
• 负责人: Alissa M Weaver
• 金额: $ 27.08万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8118809
• 关键词: Actin-Binding Protein; Actins; Adhesions; Affect; Area; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biology; Breast; Cell membrane; Cell physiology; Cells; Cellular Membrane; Cellular biology; Chemotaxis; Complex; Coupled; Couples; Coupling; Data; Discipline; F-Actin; Gene Amplification; Head and neck structure; Healed; Health; Human; In Vitro; Life; Malignant Neoplasms; Malignant Squamous Cell Neoplasm; Mechanics; Microfilaments; Molecular; Morphogenesis; Movement; Neoplasm Metastasis; Pathologic Processes; Phenotype; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipids; Photobleaching; Positioning Attribute; Proteins; Publishing; Quantitative Microscopy; Regulation; Role; Site; System; Testing; Tissues; Traction; Tube; base; cancer cell; cell motility; cofilin; depolymerization; directional cell; healing; human EMS1 protein; in vivo; mutant; neuronal cell body; outcome forecast; overexpression; polymerization; trafficking

DESCRIPTION (provided by applicant): This proposal examines mechanisms of cell motility at a fundamental level. We focus on clarifying the relationship between lamellipodial dynamics and focal complex formation, at the leading edge of a migrating cell. We are in a unique position to conduct these studies at the molecular level, because our published and preliminary data have unveiled a core mechanism whereby cortactin couples lamellipodial persistence with focal complex assembly. In Aim 1 we will test the hypothesis that this coupling is in effect akin to a "rack and pinion" steering mechanism for moving cells, which allows for a dominant lamellipodia to form and guide the cell in its direction. In Aim 2 we will define how lamellipodia persistence is generated and positively regulated. Our existing data already conclusively show that binding of cortactin to branched actin is the basis for persistence, and we will test whether cortactin stabilizes branches, super-activates Arp2/3 complex, or both. In Aim 3 we will define how persistence is spatially constrained and negatively regulated at areas of lamellipodia activity, by investigating molecular mechanisms of cortactin inhibition by phospholipids and cofilin, at the front and rear of the lamellipodia, respectively. An essential aspect of our studies is that we combine biochemistry with quantitative microscopy in order to determine not only whether molecular mechanisms can happen (e.g., in a test tube with purified components), but also whether in fact they occur in a living cell. These approaches include: chemotaxis and other cell motility assays, quantitative analyses of lamellipodial and adhesion dynamics in living cells, and careful biochemical characterization of actin binding protein mutants followed by quantification of cell phenotypes they produce. With the proposed studies on lamellipodial persistence, we are positioning ourselves at the interface of the adhesion and actin biology fields, whose integration will hopefully generate a new exciting discipline. Significance: Our studies are significant both for fundamental cell biology and human health. The molecular mechanisms we are attempting to solve are not only critical for cell motility, but also for the many cellular functions that depend on branched actin assembly, including vesicular trafficking and tissue morphogenesis. With respect to human health, these studies are particularly relevant to cancer metastasis, since cell motility is an essential component of cancer cell invasion. More directly, cortactin is well- documented to be overexpressed in a number of cancers via gene amplification, including 15% of breast and 30% of head and neck squamous cell cancer (HNSCC). Intriguingly, cortactin overexpression correlates with poor prognosis and decreased survival. Thus, the studies in this proposal are important for understanding both the fundamental regulation of dynamic branched actin assemblies and the possible role of cortactin specifically in cancer cell motility.

描述（由申请人提供）：该提案从根本上研究了细胞运动的机制。我们专注于阐明迁移细胞前缘的片状足动力​​学与焦点复合体形成之间的关系。我们处于在分子水平上进行这些研究的独特地位，因为我们已发表的初步数据揭示了皮质蛋白将片状足持久性与焦点复合物组装相结合的核心机制。在目标 1 中，我们将测试这一假设，即这种耦合实际上类似于用于移动细胞的“齿条和小齿轮”转向机制，它允许形成占主导地位的片状伪足并引导细胞朝其方向移动。在目标 2 中，我们将定义片状伪足持久性是如何产生和积极调节的。我们现有的数据已经最终表明，cortactin 与分支肌动蛋白的结合是持久性的基础，我们将测试 cortactin 是否稳定分支、超级激活 Arp2/3 复合物，或两者兼而有之。在目标 3 中，我们将通过研究磷脂和丝切蛋白分别在片状伪足的前部和后部抑制皮质素的分子机制，来定义片状伪足活动区域的持久性如何受到空间限制和负调节。我们研究的一个重要方面是，我们将生物化学与定量显微镜相结合，以便不仅确定分子机制是否可能发生（例如，在含有纯化成分的试管中），而且确定它们实际上是否发生在活细胞中。这些方法包括：趋化性和其他细胞运动测定、活细胞中板状足和粘附动力学的定量分析，以及肌动蛋白结合蛋白突变体的仔细生化表征，然后对其产生的细胞表型进行定量。通过对片状足持久性的拟议研究，我们将自己定位在粘附和肌动蛋白生物学领域的交汇处，它们的整合有望产生一个新的令人兴奋的学科。意义：我们的研究对于基础细胞生物学和人类健康都具有重要意义。我们试图解决的分子机制不仅对于细胞运动至关重要，而且对于依赖分支肌动蛋白组装的许多细胞功能也至关重要，包括囊泡运输和组织形态发生。就人类健康而言，这些研究与癌症转移特别相关，因为细胞运动是癌细胞侵袭的重要组成部分。更直接地，有充分证据表明，cortactin 通过基因扩增在多种癌症中过度表达，包括 15% 的乳腺癌和 30% 的头颈鳞状细胞癌 (HNSCC)。有趣的是，cortactin 过度表达与预后不良和生存率降低相关。因此，本提案中的研究对于理解动态分支肌动蛋白组装的基本调节以及皮质蛋白在癌细胞运动中的可能作用非常重要。

项目成果

Cortactin controls cell motility and lamellipodial dynamics by regulating ECM secretion.

• DOI: 10.1016/j.cub.2011.06.065
• 发表时间: 2011-09-13
• 期刊: Current biology : CB
• 作者: Sung BH;Zhu X;Kaverina I;Weaver AM
• 通讯作者: Weaver AM

Synthetic and tissue-derived models for studying rigidity effects on invadopodia activity.

用于研究刚性对侵袭伪足活动影响的合成和组织衍生模型。

• DOI: 10.1007/978-1-62703-538-5_10
• 发表时间: 2013
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Weaver,AlissaM;Page,JonathanM;Guelcher,ScottA;Parekh,Aron
• 通讯作者: Parekh,Aron

Regulation of cancer invasion by reactive oxygen species and Tks family scaffold proteins.

• DOI: 10.1126/scisignal.288pe56
• 发表时间: 2009-09-15
• 期刊: Science signaling
• 影响因子: 7.3
• 作者: Weaver AM

Aggressiveness of HNSCC tumors depends on expression levels of cortactin, a gene in the 11q13 amplicon.

• DOI: 10.1038/onc.2008.389
• 发表时间: 2009-01-22
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Clark, E. S.;Brown, B.;Whigham, A. S.;Kochaishvili, A.;Yarbrough, W. G.;Weaver, A. M.
• 通讯作者: Weaver, A. M.