CELL ADHESION IN TISSUE MORPHOGENESIS

组织形态发生中的细胞粘附

• 批准号: 2894793
• 负责人: WILLIAM G. CARTER
• 金额: $ 32.97万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2894793
• 关键词: cell adhesion; collagen; complementary DNA; gene induction /repression; gene mutation; human tissue; integrins; intermolecular interaction; laboratory mouse; laboratory rabbit; laminin; monoclonal antibody; oligonucleotides; polymerase chain reaction; protooncogene; site directed mutagenesis; tissue /cell culture

DESCRIPTION: Adhesion of epithelial basal cells to the basement membrane (BM) is required for maintenance of a proliferative, undifferentiated phenotype. To understand the mechanism of this adhesion requirement, we characterized ligands, receptors, and signaling proteins involved in adhesion of cultured human foreskin keratinocytes (HFKs) to the BM: (I) Laminin 5 (epiligrin) is the primary adhesive ligand in epithelial BMs of both homeostatic tissue and wound sites. It is synthesized by HFKs in wound sites. (ii) Integrin alpha-6-beta-4 in hemidesmosome-like stable anchoring contacts (SACs) and integrin alpha-3-beta-1 in focal adhesions (FAs) are adhesion receptors for laminin-5. Epithelial cells anchor to laminin 5 via alpha-6-beta-4 in homeostatic epidermis and migrate via alpha-3-beta-1 in wound sites. Integrin alpha-2-beta-1 also functions in wound repair by interacting with collagen exposed in wound sites. (iii) Phosphorylation of a tyrosine residue(s) on a new membrane-associated signaling protein, p80, is the first detectable signaling event resulting from de-attachment of HFKs from laminin 5 via alpha-6-beta-4. In contrast, ligation of alpha-2-beta-1 and alpha-3-beta-1 regulates phosphorylation of FA kinase (FAK). Synchronization of the anchorage and migratory functions to avoid interference in wound activation is regulated by receptor cross-talk. We hypothesize, that differences in cell adhesion induced by laminin 5 verses collagen results in altered phosphorylation of p80 and FAK, DNA synthesis and differentiation. We propose to: characterize p80 and its role in cell adhesion, define the mechanisms of regulatory cross-talk between the beta-1 and beta-4 receptors, and analyze the promoter of the LAMA3 gene to identify sequences responsible for transcriptional control in response to cell adhesion. these results will define the molecular mechanism through which cell adhesion regulates gene expression and wound repair in epithelium.

描述：上皮基底细胞与基底膜的粘附 (BM) 是维持增殖性、未分化的细胞所必需的 表型。 为了了解这种粘附要求的机制，我们 特征配体、受体和信号蛋白参与 培养的人包皮角质形成细胞 (HFK) 与 BM 的粘附：(I) 层粘连蛋白 5（epiligrin）是上皮 BM 中的主要粘附配体 稳态组织和伤口部位。 它是由伤口中的 HFK 合成的 网站。 (ii) 半桥粒样稳定锚定中的整合素 α-6-β-4 接触 (SAC) 和粘着斑 (FA) 中的整合素 α-3-β-1 层粘连蛋白 5 的粘附受体。 上皮细胞通过层粘连蛋白 5 锚定 表皮稳态中的 α-6-β-4 并通过 α-3-β-1 迁移 伤口部位。 整合素 α-2-β-1 也通过以下方式在伤口修复中发挥作用： 与伤口部位暴露的胶原蛋白相互作用。 (iii) 磷酸化 新的膜相关信号蛋白 p80 上的酪氨酸残基， 是由 HFK 脱离引起的第一个可检测到的信号事件 来自层粘连蛋白 5 通过 alpha-6-beta-4。 相反，α-2-β-1 的连接 α-3-β-1 调节 FA 激酶 (FAK) 的磷酸化。 锚地和迁移功能的同步以避免 伤口激活的干扰是由受体串扰调节的。 我们 假设层粘连蛋白 5 诗节诱导的细胞粘附差异 胶原蛋白导致 p80 和 FAK 磷酸化、DNA 合成发生改变 和差异化。 我们建议：表征 p80 及其在细胞中的作用 粘附，定义 beta-1 之间的调控串扰机制 和β-4受体，并分析LAMA3基因的启动子以鉴定 负责响应细胞转录控制的序列 附着力。 这些结果将定义分子机制 细胞粘附调节上皮细胞的基因表达和伤口修复。